Result

Search results for the GEO ID: GSE28441

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Title

Source name

Description

Characteristics

GSM703090

GPL85

Sorted TRH, GFP+, biological rep 1

TRH-GFP Transfected Hypothalamic Neurons, Sorted GFP+

cell type: TRH-GFP Transfected Hypothalamic Neurons, Sorted GFP+ tissue: Hypothalamus strain: Wistar developmental stage: Embryo age: E17

Primary cultures of E17 hypotalamic neurons were transfected with a GFP TRH-promoter drived vector. The culture was sorted afterwards in order to obtain only GFP positive cells

Sample_geo_accession	GSM703090
Sample_status	Public on May 27 2011
Sample_submission_date	Apr 07 2011
Sample_last_update_date	May 27 2011
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Rattus norvegicus
Sample_taxid_ch1	10116
Sample_treatment_protocol_ch1	Twenty-four hours after seeding, cells were transfected essentially as described before. In general, 8 mg of branched polyethylenimine (PEI) (600-1000 KDa; Fluka) solution was diluted in 10 ml of water, pH adjusted to 6.9 with 0.2 N HCl and the solution filtered (Millipore, 0.22 µm). PEI (30 µl) and plasmid DNA (10 µg) were separately diluted to adjust NaCl to 150 mM in a final volume of 50 µl, vortexed and incubated for 10 min at room temperature; subsequently, the polymer solution was added to the DNA, vortexed-mixed, incubated for 10 min at room temperature followed by the addition of 900 µl of serum-free DMEM. The supplemented DMEM was removed from the culture dishes and the transfection mixture was added. After 3 hours incubation, transfection mix was removed and fresh supplemented DMEM was added.
Sample_growth_protocol_ch1	Animals used were Wistar rats raised at the animal facility of the Instituto de Biotecnología, UNAM. Animals were maintained in standard environmental conditions (lights on between 0700-1900 h, temperature 21 ± 2 oC) and received rat chow and tap water ad libitum. Animal care and protocols were approved by the Animal Care and Ethics Committee of the Institute following the guidelines for the use of animals in neuroscience research of the Society for Neuroscience, USA.
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Total RNA (10 µg) was extracted from three different cell populations: i) sorted TRH-GFP+ cells (GFP+); ii) TRH-GFP+ and GFP- mixed cells (GFP+/-) passed through the FACS but not sorted, and iii) non transfected cells (NT). To obtain a sufficient amount of RNA for each cell population, the number of independent experiments pooled for the GFP+ sample was higher than for the other samples. Therefore, a pool of six independent experiments was used to prepare total RNA from the GFP+ and three independent experiments for GFP+/- or NT cells.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using SuperScript II Reverse Transcriptase, RNase H, and DNA polymerase (Invitrogen). After precipitation with 5M NH4OAc and ethanol, the cDNA was used as a template in a biotin-labeled in vitro transcription reaction (Enzo BioArray, Affymetrix).
Sample_hyb_protocol	Resulting target cRNA was collected on RNAeasy columns (QIAGEN, Valencia, CA) and then fragmented for hybridization on the microarrays. Biotinylated target cRNA (15 µg) was hybridized to the array and then processed using the Affymetrix GeneChip Fluidics Workstation 400
Sample_scan_protocol	After binding with phycoerythrin-coupled avidin, microarrays were scanned on a Hewlett-Packard Gene Array Scanner (Hewlett-Packard Co., Palo Alto, CA).
Sample_data_processing	Results were analyzed using Affymetrix MAS 5.0 software. Individual microarrays were scaled to produce a mean signal intensity of 125. Iterative comparisons of different microarray datasets were done with MAS 5.0 comparison analysis.
Sample_platform_id	GPL85
Sample_contact_name	Leonor,,Perez-Martinez
Sample_contact_email	leonor@ibt.unam.mx
Sample_contact_phone	52-777-329-1600
Sample_contact_fax	52-777-317-2388
Sample_contact_laboratory	Neuroimmunobiology
Sample_contact_department	Medicina Molecular y Bioprocesos
Sample_contact_institute	University of Mexico
Sample_contact_address	Avenida Universidad 2001
Sample_contact_city	Cuernavaca
Sample_contact_state	Morelos
Sample_contact_zip/postal_code	62271
Sample_contact_country	Mexico
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703090/suppl/GSM703090.CEL.gz
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703090/suppl/GSM703090.CHP.gz
Sample_series_id	GSE28441
Sample_data_row_count	8799

GSM703091

GPL85

Sorted TRH, GFP+, biological rep 2

TRH-GFP Transfected Hypothalamic Neurons, Sorted GFP+

cell type: TRH-GFP Transfected Hypothalamic Neurons, Sorted GFP+ tissue: Hypothalamus strain: Wistar developmental stage: Embryo age: E17

Primary cultures of E17 hypotalamic neurons were transfected with a GFP TRH-promoter drived vector. The culture was sorted afterwards in order to obtain only GFP positive cells

Sample_geo_accession	GSM703091
Sample_status	Public on May 27 2011
Sample_submission_date	Apr 07 2011
Sample_last_update_date	May 27 2011
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Rattus norvegicus
Sample_taxid_ch1	10116
Sample_treatment_protocol_ch1	Twenty-four hours after seeding, cells were transfected essentially as described before. In general, 8 mg of branched polyethylenimine (PEI) (600-1000 KDa; Fluka) solution was diluted in 10 ml of water, pH adjusted to 6.9 with 0.2 N HCl and the solution filtered (Millipore, 0.22 µm). PEI (30 µl) and plasmid DNA (10 µg) were separately diluted to adjust NaCl to 150 mM in a final volume of 50 µl, vortexed and incubated for 10 min at room temperature; subsequently, the polymer solution was added to the DNA, vortexed-mixed, incubated for 10 min at room temperature followed by the addition of 900 µl of serum-free DMEM. The supplemented DMEM was removed from the culture dishes and the transfection mixture was added. After 3 hours incubation, transfection mix was removed and fresh supplemented DMEM was added.
Sample_growth_protocol_ch1	Animals used were Wistar rats raised at the animal facility of the Instituto de Biotecnología, UNAM. Animals were maintained in standard environmental conditions (lights on between 0700-1900 h, temperature 21 ± 2 oC) and received rat chow and tap water ad libitum. Animal care and protocols were approved by the Animal Care and Ethics Committee of the Institute following the guidelines for the use of animals in neuroscience research of the Society for Neuroscience, USA.
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Total RNA (10 µg) was extracted from three different cell populations: i) sorted TRH-GFP+ cells (GFP+); ii) TRH-GFP+ and GFP- mixed cells (GFP+/-) passed through the FACS but not sorted, and iii) non transfected cells (NT). To obtain a sufficient amount of RNA for each cell population, the number of independent experiments pooled for the GFP+ sample was higher than for the other samples. Therefore, a pool of six independent experiments was used to prepare total RNA from the GFP+ and three independent experiments for GFP+/- or NT cells.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using SuperScript II Reverse Transcriptase, RNase H, and DNA polymerase (Invitrogen). After precipitation with 5M NH4OAc and ethanol, the cDNA was used as a template in a biotin-labeled in vitro transcription reaction (Enzo BioArray, Affymetrix).
Sample_hyb_protocol	Resulting target cRNA was collected on RNAeasy columns (QIAGEN, Valencia, CA) and then fragmented for hybridization on the microarrays. Biotinylated target cRNA (15 µg) was hybridized to the array and then processed using the Affymetrix GeneChip Fluidics Workstation 400
Sample_scan_protocol	After binding with phycoerythrin-coupled avidin, microarrays were scanned on a Hewlett-Packard Gene Array Scanner (Hewlett-Packard Co., Palo Alto, CA).
Sample_data_processing	Results were analyzed using Affymetrix MAS 5.0 software. Individual microarrays were scaled to produce a mean signal intensity of 125. Iterative comparisons of different microarray datasets were done with MAS 5.0 comparison analysis.
Sample_platform_id	GPL85
Sample_contact_name	Leonor,,Perez-Martinez
Sample_contact_email	leonor@ibt.unam.mx
Sample_contact_phone	52-777-329-1600
Sample_contact_fax	52-777-317-2388
Sample_contact_laboratory	Neuroimmunobiology
Sample_contact_department	Medicina Molecular y Bioprocesos
Sample_contact_institute	University of Mexico
Sample_contact_address	Avenida Universidad 2001
Sample_contact_city	Cuernavaca
Sample_contact_state	Morelos
Sample_contact_zip/postal_code	62271
Sample_contact_country	Mexico
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703091/suppl/GSM703091.CEL.gz
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703091/suppl/GSM703091.CHP.gz
Sample_series_id	GSE28441
Sample_data_row_count	8799

GSM703092

GPL85

Total Cell Population, GFP +/-, biological rep 1

TRH-GFP Transfected Hypothalamic Neurons, Total Population GFP+/-

cell type: TRH-GFP Transfected Hypothalamic Neurons, Total Population GFP+/- tissue: Hypothalamus strain: Wistar developmental stage: Embryo age: E17

Primary cultures of E17 hypotalamic neurons were transfected with a GFP TRH-promoter drived vector. RNA from total population (GFP+/- cells) was used

Sample_geo_accession	GSM703092
Sample_status	Public on May 27 2011
Sample_submission_date	Apr 07 2011
Sample_last_update_date	May 27 2011
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Rattus norvegicus
Sample_taxid_ch1	10116
Sample_treatment_protocol_ch1	Twenty-four hours after seeding, cells were transfected essentially as described before. In general, 8 mg of branched polyethylenimine (PEI) (600-1000 KDa; Fluka) solution was diluted in 10 ml of water, pH adjusted to 6.9 with 0.2 N HCl and the solution filtered (Millipore, 0.22 µm). PEI (30 µl) and plasmid DNA (10 µg) were separately diluted to adjust NaCl to 150 mM in a final volume of 50 µl, vortexed and incubated for 10 min at room temperature; subsequently, the polymer solution was added to the DNA, vortexed-mixed, incubated for 10 min at room temperature followed by the addition of 900 µl of serum-free DMEM. The supplemented DMEM was removed from the culture dishes and the transfection mixture was added. After 3 hours incubation, transfection mix was removed and fresh supplemented DMEM was added.
Sample_growth_protocol_ch1	Animals used were Wistar rats raised at the animal facility of the Instituto de Biotecnología, UNAM. Animals were maintained in standard environmental conditions (lights on between 0700-1900 h, temperature 21 ± 2 oC) and received rat chow and tap water ad libitum. Animal care and protocols were approved by the Animal Care and Ethics Committee of the Institute following the guidelines for the use of animals in neuroscience research of the Society for Neuroscience, USA.
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Total RNA (10 µg) was extracted from three different cell populations: i) sorted TRH-GFP+ cells (GFP+); ii) TRH-GFP+ and GFP- mixed cells (GFP+/-) passed through the FACS but not sorted, and iii) non transfected cells (NT). To obtain a sufficient amount of RNA for each cell population, the number of independent experiments pooled for the GFP+ sample was higher than for the other samples. Therefore, a pool of six independent experiments was used to prepare total RNA from the GFP+ and three independent experiments for GFP+/- or NT cells.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using SuperScript II Reverse Transcriptase, RNase H, and DNA polymerase (Invitrogen). After precipitation with 5M NH4OAc and ethanol, the cDNA was used as a template in a biotin-labeled in vitro transcription reaction (Enzo BioArray, Affymetrix).
Sample_hyb_protocol	Resulting target cRNA was collected on RNAeasy columns (QIAGEN, Valencia, CA) and then fragmented for hybridization on the microarrays. Biotinylated target cRNA (15 µg) was hybridized to the array and then processed using the Affymetrix GeneChip Fluidics Workstation 400
Sample_scan_protocol	After binding with phycoerythrin-coupled avidin, microarrays were scanned on a Hewlett-Packard Gene Array Scanner (Hewlett-Packard Co., Palo Alto, CA).
Sample_data_processing	Results were analyzed using Affymetrix MAS 5.0 software. Individual microarrays were scaled to produce a mean signal intensity of 125. Iterative comparisons of different microarray datasets were done with MAS 5.0 comparison analysis.
Sample_platform_id	GPL85
Sample_contact_name	Leonor,,Perez-Martinez
Sample_contact_email	leonor@ibt.unam.mx
Sample_contact_phone	52-777-329-1600
Sample_contact_fax	52-777-317-2388
Sample_contact_laboratory	Neuroimmunobiology
Sample_contact_department	Medicina Molecular y Bioprocesos
Sample_contact_institute	University of Mexico
Sample_contact_address	Avenida Universidad 2001
Sample_contact_city	Cuernavaca
Sample_contact_state	Morelos
Sample_contact_zip/postal_code	62271
Sample_contact_country	Mexico
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703092/suppl/GSM703092.CEL.gz
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703092/suppl/GSM703092.CHP.gz
Sample_series_id	GSE28441
Sample_data_row_count	8799

GSM703093

GPL85

Total Cell Population, GFP +/-, biological rep 2

TRH-GFP Transfected Hypothalamic Neurons, Total Population GFP+/-

cell type: TRH-GFP Transfected Hypothalamic Neurons, Total Population GFP+/- tissue: Hypothalamus strain: Wistar developmental stage: Embryo age: E17

Primary cultures of E17 hypotalamic neurons were transfected with a GFP TRH-promoter drived vector. RNA from total population (GFP+/- cells) was used

Sample_geo_accession	GSM703093
Sample_status	Public on May 27 2011
Sample_submission_date	Apr 07 2011
Sample_last_update_date	May 27 2011
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Rattus norvegicus
Sample_taxid_ch1	10116
Sample_treatment_protocol_ch1	Twenty-four hours after seeding, cells were transfected essentially as described before. In general, 8 mg of branched polyethylenimine (PEI) (600-1000 KDa; Fluka) solution was diluted in 10 ml of water, pH adjusted to 6.9 with 0.2 N HCl and the solution filtered (Millipore, 0.22 µm). PEI (30 µl) and plasmid DNA (10 µg) were separately diluted to adjust NaCl to 150 mM in a final volume of 50 µl, vortexed and incubated for 10 min at room temperature; subsequently, the polymer solution was added to the DNA, vortexed-mixed, incubated for 10 min at room temperature followed by the addition of 900 µl of serum-free DMEM. The supplemented DMEM was removed from the culture dishes and the transfection mixture was added. After 3 hours incubation, transfection mix was removed and fresh supplemented DMEM was added.
Sample_growth_protocol_ch1	Animals used were Wistar rats raised at the animal facility of the Instituto de Biotecnología, UNAM. Animals were maintained in standard environmental conditions (lights on between 0700-1900 h, temperature 21 ± 2 oC) and received rat chow and tap water ad libitum. Animal care and protocols were approved by the Animal Care and Ethics Committee of the Institute following the guidelines for the use of animals in neuroscience research of the Society for Neuroscience, USA.
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Total RNA (10 µg) was extracted from three different cell populations: i) sorted TRH-GFP+ cells (GFP+); ii) TRH-GFP+ and GFP- mixed cells (GFP+/-) passed through the FACS but not sorted, and iii) non transfected cells (NT). To obtain a sufficient amount of RNA for each cell population, the number of independent experiments pooled for the GFP+ sample was higher than for the other samples. Therefore, a pool of six independent experiments was used to prepare total RNA from the GFP+ and three independent experiments for GFP+/- or NT cells.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using SuperScript II Reverse Transcriptase, RNase H, and DNA polymerase (Invitrogen). After precipitation with 5M NH4OAc and ethanol, the cDNA was used as a template in a biotin-labeled in vitro transcription reaction (Enzo BioArray, Affymetrix).
Sample_hyb_protocol	Resulting target cRNA was collected on RNAeasy columns (QIAGEN, Valencia, CA) and then fragmented for hybridization on the microarrays. Biotinylated target cRNA (15 µg) was hybridized to the array and then processed using the Affymetrix GeneChip Fluidics Workstation 400
Sample_scan_protocol	After binding with phycoerythrin-coupled avidin, microarrays were scanned on a Hewlett-Packard Gene Array Scanner (Hewlett-Packard Co., Palo Alto, CA).
Sample_data_processing	Results were analyzed using Affymetrix MAS 5.0 software. Individual microarrays were scaled to produce a mean signal intensity of 125. Iterative comparisons of different microarray datasets were done with MAS 5.0 comparison analysis.
Sample_platform_id	GPL85
Sample_contact_name	Leonor,,Perez-Martinez
Sample_contact_email	leonor@ibt.unam.mx
Sample_contact_phone	52-777-329-1600
Sample_contact_fax	52-777-317-2388
Sample_contact_laboratory	Neuroimmunobiology
Sample_contact_department	Medicina Molecular y Bioprocesos
Sample_contact_institute	University of Mexico
Sample_contact_address	Avenida Universidad 2001
Sample_contact_city	Cuernavaca
Sample_contact_state	Morelos
Sample_contact_zip/postal_code	62271
Sample_contact_country	Mexico
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703093/suppl/GSM703093.CEL.gz
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703093/suppl/GSM703093.CHP.gz
Sample_series_id	GSE28441
Sample_data_row_count	8799

GSM703094

GPL85

Non transfected Cells

Non-transfected Hypothalamic Neurons

cell type: Non-transfected Hypothalamic Neurons tissue: Hypothalamus strain: Wistar developmental stage: Embryo age: E17

Primary cultures of E17 hypotalamic neurons.

Sample_geo_accession	GSM703094
Sample_status	Public on May 27 2011
Sample_submission_date	Apr 07 2011
Sample_last_update_date	May 27 2011
Sample_type	RNA
Sample_channel_count	1
Sample_organism_ch1	Rattus norvegicus
Sample_taxid_ch1	10116
Sample_treatment_protocol_ch1	Twenty-four hours after seeding, cells were transfected essentially as described before. In general, 8 mg of branched polyethylenimine (PEI) (600-1000 KDa; Fluka) solution was diluted in 10 ml of water, pH adjusted to 6.9 with 0.2 N HCl and the solution filtered (Millipore, 0.22 µm). PEI (30 µl) and plasmid DNA (10 µg) were separately diluted to adjust NaCl to 150 mM in a final volume of 50 µl, vortexed and incubated for 10 min at room temperature; subsequently, the polymer solution was added to the DNA, vortexed-mixed, incubated for 10 min at room temperature followed by the addition of 900 µl of serum-free DMEM. The supplemented DMEM was removed from the culture dishes and the transfection mixture was added. After 3 hours incubation, transfection mix was removed and fresh supplemented DMEM was added.
Sample_growth_protocol_ch1	Animals used were Wistar rats raised at the animal facility of the Instituto de Biotecnología, UNAM. Animals were maintained in standard environmental conditions (lights on between 0700-1900 h, temperature 21 ± 2 oC) and received rat chow and tap water ad libitum. Animal care and protocols were approved by the Animal Care and Ethics Committee of the Institute following the guidelines for the use of animals in neuroscience research of the Society for Neuroscience, USA.
Sample_molecule_ch1	total RNA
Sample_extract_protocol_ch1	Total RNA (10 µg) was extracted from three different cell populations: i) sorted TRH-GFP+ cells (GFP+); ii) TRH-GFP+ and GFP- mixed cells (GFP+/-) passed through the FACS but not sorted, and iii) non transfected cells (NT). To obtain a sufficient amount of RNA for each cell population, the number of independent experiments pooled for the GFP+ sample was higher than for the other samples. Therefore, a pool of six independent experiments was used to prepare total RNA from the GFP+ and three independent experiments for GFP+/- or NT cells.
Sample_label_ch1	Biotin
Sample_label_protocol_ch1	RNA was reverse-transcribed into double-stranded cDNA with a T7 promoter-containing primer using SuperScript II Reverse Transcriptase, RNase H, and DNA polymerase (Invitrogen). After precipitation with 5M NH4OAc and ethanol, the cDNA was used as a template in a biotin-labeled in vitro transcription reaction (Enzo BioArray, Affymetrix).
Sample_hyb_protocol	Resulting target cRNA was collected on RNAeasy columns (QIAGEN, Valencia, CA) and then fragmented for hybridization on the microarrays. Biotinylated target cRNA (15 µg) was hybridized to the array and then processed using the Affymetrix GeneChip Fluidics Workstation 400
Sample_scan_protocol	After binding with phycoerythrin-coupled avidin, microarrays were scanned on a Hewlett-Packard Gene Array Scanner (Hewlett-Packard Co., Palo Alto, CA).
Sample_data_processing	Results were analyzed using Affymetrix MAS 5.0 software. Individual microarrays were scaled to produce a mean signal intensity of 125. Iterative comparisons of different microarray datasets were done with MAS 5.0 comparison analysis.
Sample_platform_id	GPL85
Sample_contact_name	Leonor,,Perez-Martinez
Sample_contact_email	leonor@ibt.unam.mx
Sample_contact_phone	52-777-329-1600
Sample_contact_fax	52-777-317-2388
Sample_contact_laboratory	Neuroimmunobiology
Sample_contact_department	Medicina Molecular y Bioprocesos
Sample_contact_institute	University of Mexico
Sample_contact_address	Avenida Universidad 2001
Sample_contact_city	Cuernavaca
Sample_contact_state	Morelos
Sample_contact_zip/postal_code	62271
Sample_contact_country	Mexico
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703094/suppl/GSM703094.CEL.gz
Sample_supplementary_file	ftp://ftp.ncbi.nlm.nih.gov/geo/samples/GSM703nnn/GSM703094/suppl/GSM703094.CHP.gz
Sample_series_id	GSE28441
Sample_data_row_count	8799

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