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Status |
Public on May 15, 2024 |
Title |
RNA-seq analysis of B cells cultured with LPS and IL-12 and/or IFN gamma |
Organism |
Mus musculus |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
Purpose: We aimed to find genes differentially regulated by IL-12 and IFNg alone, and in combination, over 4 and 24 hours of stimulation. Methods : Spleens were processed and B cell cultures prepared, in triplicate, according to the manuscript. After the indicated times, cells were washed in PBS, resuspended in 350 ul RLT Plus Buffer (Qiagen) with 1% 2-Me, and frozen at -80 ⁰C. Results: At 4 hours, the main effect of IL-12 is to induce IFNg production from B cells, whereas IFNg induces many gene changes. Comparing 24 hours to the 4 hour stimulation, IFNg alone was insufficient to maintain the majority of IFNg-induced 4 hour changes, while adding IL-12 sustained a portion of the 4 hour gene changes. Additionally, in combination IL-12 and IFNg cooperate to induce unique gene changes that are not shared with either the individual cytokines at either timepoint.
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Overall design |
We used recombinant IL-12 and IFNg or anti-IL-12 and anti-IFNg blocking antibodies to assess the contribution of each cytokine alone and in combination.
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Contributor(s) |
Elsner RA, Smita S, Shlomchik MJ |
Citation(s) |
38862796 |
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Submission date |
May 08, 2023 |
Last update date |
Jul 01, 2024 |
Contact name |
Mark Shlomchik |
E-mail(s) |
shlomchiklab@gmail.com
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Organization name |
University of Pittsburgh
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Department |
Department of Immunology
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Street address |
The Assembly, 5051 Centre Avenue
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City |
Pittsburgh |
State/province |
PA |
ZIP/Postal code |
15213 |
Country |
USA |
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Platforms (1) |
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Samples (42)
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GSM7306190 |
4 hours, IL-12 at 2000 pg/mL, IFNg block, mouse 1 |
GSM7306191 |
4 hours, IL-12 at 2000 pg/mL, IFNg block, mouse 2 |
GSM7306192 |
4 hours, IL-12 at 2000 pg/mL, IFNg block, mouse 3 |
GSM7306193 |
4 hours, IL-12 at 2000 pg/mL, IFNg at 0.5 ng/mL, mouse 1 |
GSM7306194 |
4 hours, IL-12 at 2000 pg/mL, IFNg at 0.5 ng/mL, mouse 2 |
GSM7306195 |
4 hours, IL-12 at 2000 pg/mL, IFNg at 0.5 ng/mL, mouse 3 |
GSM7306196 |
4 hours, IL-12 at 200 pg/mL, IFNg at 0.5 ng/mL, mouse 1 |
GSM7306197 |
4 hours, IL-12 at 200 pg/mL, IFNg at 0.5 ng/mL, mouse 2 |
GSM7306198 |
4 hours, IL-12 at 200 pg/mL, IFNg at 0.5 ng/mL, mouse 3 |
GSM7306199 |
4 hours, IL-12 at 200 pg/mL, IFNg block, mouse 1 |
GSM7306200 |
4 hours, IL-12 at 200 pg/mL, IFNg block, mouse 2 |
GSM7306201 |
4 hours, IL-12 at 200 pg/mL, IFNg block, mouse 3 |
GSM7306202 |
4 hours, IL-12 at 20 pg/mL, IFNg at 0.5 ng/mL, mouse 1 |
GSM7306203 |
4 hours, IL-12 at 20 pg/mL, IFNg at 0.5 ng/mL, mouse 2 |
GSM7306204 |
4 hours, IL-12 at 20 pg/mL, IFNg at 0.5 ng/mL, mouse 3 |
GSM7306205 |
4 hours, IL-12 at 20 pg/mL, IFNg block, mouse 1 |
GSM7306206 |
4 hours, IL-12 at 20 pg/mL, IFNg block, mouse 2 |
GSM7306207 |
4 hours, IL-12 at 20 pg/mL, IFNg block, mouse 3 |
GSM7306208 |
24 hours, IL-12 at 20 pg/mL, IFNg at 0.5 ng/mL, mouse 1 |
GSM7306209 |
24 hours, IL-12 at 20 pg/mL, IFNg at 0.5 ng/mL, mouse 2 |
GSM7306210 |
24 hours, IL-12 at 20 pg/mL, IFNg at 0.5 ng/mL, mouse 3 |
GSM7306211 |
24 hours, IL-12 at 20 pg/mL, IFNg block, mouse 1 |
GSM7306212 |
24 hours, IL-12 at 20 pg/mL, IFNg block, mouse 2 |
GSM7306213 |
24 hours, IL-12 at 20 pg/mL, IFNg block, mouse 3 |
GSM7306214 |
24 hours, IL-12 block, IFNg at 0.5 ng/mL, mouse 1 |
GSM7306215 |
24 hours, IL-12 block, IFNg at 0.5 ng/mL, mouse 2 |
GSM7306216 |
24 hours, IL-12 block, IFNg at 0.5 ng/mL, mouse 3 |
GSM7306217 |
24 hours, IL-12 block, IFNg block, mouse 1 |
GSM7306218 |
24 hours, IL-12 block, IFNg block, mouse 2 |
GSM7306219 |
24 hours, IL-12 block, IFNg block, mouse 3 |
GSM7306220 |
4 hours, IL-12 block, IFNg at 0.5 ng/mL, mouse 1 |
GSM7306221 |
4 hours, IL-12 block, IFNg at 0.5 ng/mL, mouse 2 |
GSM7306222 |
4 hours, IL-12 block, IFNg at 0.5 ng/mL, mouse 3 |
GSM7306223 |
4 hours, IL-12 block, IFNg at 3 ng/mL, mouse 1 |
GSM7306224 |
4 hours, IL-12 block, IFNg at 3 ng/mL, mouse 2 |
GSM7306225 |
4 hours, IL-12 block, IFNg at 3 ng/mL, mouse 3 |
GSM7306226 |
4 hours, IL-12 block, IFNg block, mouse 1 |
GSM7306227 |
4 hours, IL-12 block, IFNg block, mouse 2 |
GSM7306228 |
4 hours, IL-12 block, IFNg block, mouse 3 |
GSM7306229 |
4 hours, no added IL-12 nor blocked, IFNg at 3 ng/mL, mouse 1 |
GSM7306230 |
4 hours, no added IL-12 nor blocked, IFNg at 3 ng/mL, mouse 2 |
GSM7306231 |
4 hours, no added IL-12 nor blocked, IFNg at 3 ng/mL, mouse 3 |
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Relations |
BioProject |
PRJNA970259 |
Supplementary file |
Size |
Download |
File type/resource |
GSE231931_RAW.tar |
3.0 Mb |
(http)(custom) |
TAR (of TXT) |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
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