Journal: medRxiv

Article Title: Allelic effects on KLHL17 expression likely mediated by JunB/D underlie a PDAC GWAS signal at chr1p36.33

doi: 10.1101/2024.09.16.24313748

Figure Lengend Snippet: a) in silico TF binding predictions; b) EMSA using TPA-stimulated nuclear HeLa extract and fluorescently labeled oligonucleotide. Arrow indicates the allele-preferential binding; c) Luciferase reporter assay using DMSO and TPA stimulation and the rs13303160 sequence as an enhancer in the PANC-1 cell line; luciferase activity reported relative to the Empty Vector (EV). Unpaired t-tests were performed on the relative luciferase activity of the A/G ratio compared to A/A; d) Representative EMSAs with increasing amounts of recombinant Fos proteins (from left to right: c-Fos; FosB; Fos1L; Fos2L); e) Representative EMSAs with increasing amounts of recombinant Jun proteins (from left to right: c-Jun, JunB, JunD). Arrow indicates the allele-specific binding; f, g) Supershift EMSA with antibodies against JunB and JunD respectively using both TPA-stimulated nuclear lysate and recombinant protein; Arrows denote the shift in the bands; h) ChIP-qPCR in SW1990 PDAC cells for JunB and JunD using 3 primer sets (PS) surrounding the SNP. Positive controls are from a JunB ChIP-seq performed in the CFPAC1 PDAC cell line. Negative control is from a quiescent region on 1p36.33; i) TaqMan genotyping assay for rs13303160 using immunoprecipitated DNA from the ChIP. The ratio of A to G was determined relative to the quantity of A and G alleles in the input DNA. For all graphs, error bars represent the SEM. Unpaired two-tailed t-tests were performed; * P <0.05; ** P <0.01; *** P <0.001.

Article Snippet: Recombinant ELF1 (TP760629), ELF2 (TP760288), ELF3 (TP300631), ELF4 (TP761826), JUNB (TP303595), JUND (TP316958 4), c-FOS (TP760257), FOS1L (TP302104), FOSB (TP762032), FOS2L (TP760114) protein were purchased from Origene (Rockville, MD). c-JUN was purchased from Abcam (Waltham, MA) (ab84134).

Techniques: In Silico, Binding Assay, Labeling, Luciferase, Reporter Assay, Sequencing, Activity Assay, Plasmid Preparation, Recombinant, ChIP-sequencing, Negative Control, Genotyping Assay, Immunoprecipitation, Two Tailed Test

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Rpkm values are plotted for FOSL1 [left] and FOSL2 [right] RNA at different time points of activation (Th0) or Th17-polarization, using published RNA-seq data ( Oncotarget ). ( B ) Immunoblot images (lower panel) show FOSL1 [left] and FOSL2 [right] protein levels in Th0 and Th17-polarizing cells, over a time-course. Actin was used as loading control. Blots from three biological replicates were quantified using ImageJ and the corresponding FOSL intensity values (normalized to actin) are plotted as a line graph in the above panel. ( C ) Flow cytometry analysis of FOSL1 [left] and FOSL2 [right] expression in naive CD4+ T cells cultured for 24h, under conditions of activation (Th0), Th17-polarization, or activation in presence of the Th17-cytokines (used either alone or in combination). Bar plot shows median fluorescence intensity (MFI) values normalized to Th0, for three biological replicates. Statistical significance was calculated by comparing each condition to Th0. ( D ) Flow cytometry analysis of FOSL1 [left] and FOSL2 [right] protein levels in non-targeting (SCR) versus STAT3 KD Th17 cells, at 72h of polarization. Graph shows MFI values normalized to SCR for four biological replicates. ( E ) Naive CD4 + T cells were silenced for FOSL1 [left] or FOSL2 [right] using two different siRNAs each, and further polarized to Th17-fate for 24h. Knockdown was analyzed using immunoblotting. Representative blots for three biological replicates are shown. ( F ) ELISA was used to estimate IL-17A secretion in supernatants of FOSL1 [left] and FOSL2-silenced [right] Th17 cells, at 72h of polarization. Values were first normalized to live cell count, followed by normalization with SCR. Data represents four or five biological replicates, as indicated. Graphs in the above panels show mean ± standard error of the mean (SEM). Statistical significance is calculated using two-tailed Student’s t test (*p < 0.05; **p < 0.01, ***p < 0.001).

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Activation Assay, RNA Sequencing Assay, Western Blot, Flow Cytometry, Expressing, Cell Culture, Fluorescence, Enzyme-linked Immunosorbent Assay, Cell Counting, Two Tailed Test

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Immunoblot images show FOSL1 (above) and FOSL2 (below) protein levels in naive CD4 + T cells cultured under activation (Th0) or Th17 differentiation conditions, for the indicated time points. Actin has been used as loading control. Data represents biological replicates for . ( B ) UCSC genome browser snapshots indicate the binding of STAT3 over the promoter of FOSL2 (above panel) and not FOSL1 (below panel), in Th17 cells cultured for 0.5 h and 72h. Figures were derived using bed files of STAT3 ChIP-seq data from Tripathi et al., 2017 Cell Reports. ( C ) Nucleofection workflow for FOSL1/FOSL2 knockdown (KD). Naive CD4 + T cells were treated with FOSL1- or FOSL2-targeting siRNAs, rested for 36-40 h, and further cultured under Th17-polarizing conditions (IL-6, IL-1β and TGF-β) for 72h. ( D ) Immunoblots depict FOSL1 and FOSL2 protein levels in naive CD4 + T cells that were silenced for the respective factors and cultured under Th17-polarizing conditions for 24h. Non-targeting siRNA (Scramble or SCR) was used as nucleofection control and actin was used as loading control. Blots shown are biological replicates for .

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Western Blot, Cell Culture, Activation Assay, Binding Assay, Derivative Assay, ChIP-sequencing

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Naive CD4 + T cells were silenced for FOSL1, FOSL2 or both factors in parallel (double KD; DKD), and cultured under Th17-polarizing conditions for 24h. Total FOSL1 [left] or FOSL2 [right] protein was stained (Alexa-647) and analysed by flow cytometry. Non-targeting siRNA (SCR) was used as nucleofection control. Representative histograms for four biological replicates are shown. ( B ) Naive CD4 + T cells were treated with in-vitro transcribed GFP-FOSL1 RNA, GFP-FOSL2 RNA or both (double OE; DOE). After resting the cells for 18–20h, total FOSL1 [left] or FOSL2 [right] protein was stained (Alexa-647) and analysed by flow cytometry. GFP RNA were used as nucleofection control. Representative histograms for four biological replicates are shown. ( C and D ) Bar plot shows ELISA results for secreted IL-17A levels in supernatants of FOSL KD/DKD (panel C) or FOSL OE/DOE Th17 cells (panel D), at 72h of polarization. Values were first normalized to live cell count, and then to the respective control condition (SCR or GFP). Data represent four biological replicates. ( E and F ) qRT-PCR analysis for measurement of IL-17A [left] and IL-17F [right] RNA levels in FOSL KD/DKD (panel E) or FOSL OE/DOE Th17 cells (panel F), at 72h of polarization. Fold-change normalized to the respective controls (SCR or empty GFP) was plotted for four biological replicates. For panels C-F, plots show mean ± SEM. Statistical significance is calculated using two-tailed Student’s t test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p<0.0001).

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Cell Culture, Staining, Flow Cytometry, In Vitro, Enzyme-linked Immunosorbent Assay, Cell Counting, Quantitative RT-PCR, Two Tailed Test

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A and ) FOSL KD/DKD (panel A) and FOSL OE/DOE (panel B) Th17 cells were labelled (Alexa-647) for total FOSL1 and FOSL2 protein at 24h of polarization. Expression of the corresponding factors was analyzed using flow cytometry and overlay histograms were plotted [FOSL1, left; FOSL2, right]. Figure shows biological replicates for and .

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A and B ) Heatmap in panel A shows the DE genes that are more profoundly altered in FOSL DKD Th17 cells relative to the single KD controls, at 24h (above) and 72h (below) of polarization. Panel B includes the DE genes that show enhanced changes in FOSL DOE Th17 cells as compared to the single OE controls at 72h of polarization. Genes with Th17-relevance are highlighted; upregulated genes are in red, and downregulated ones are in blue. Log2fold-change was calculated relative to the respective control conditions (i.e., SCR or GFP). ( C and D ) Ingenuity pathway analysis was used to identify signaling pathways that are altered upon FOSL DKD (panel C) or DOE (panel D). ( E and F) Genome-wide expression analysis of FOSL DKD and FOSL DOE Th17 cells. Volcano plots in Panel E highlight the Th17-associated transcripts that are differentially expressed upon co-depletion of FOSL1 and FOSL2, at 24h [left] and 72h [right] of Th17 polarization. Panel F highlights the Th17-associated genes that are differentially expressed upon parallel over-expression of FOSL1 and FOSL2, at 72h of Th17 polarization. Targets with FDR ≤ 0.1 and |fold-change| ≥ 1.8 have been plotted. Upregulated genes are in red, and the downregulated ones are in blue. ( G ) Heatmap depicts the DE genes that show opposite expression changes in FOSL DKD versus DOE conditions, at the indicated time points of Th17 polarization. Th17-relevant genes have been highlighted.

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Genome Wide, Expressing, Over Expression

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Immunofluorescence images showing nuclear localization of FOSL1 (red, above panel) and FOSL2 (red, below panel) in Th17 cells polarized for 72h. Lamin A/C (in green) marks the nuclear periphery, whereas phalloidin (in blue) stains the cytoplasmic actin. ( B ) ChIP-seq analysis was performed for FOSL1 and FOSL2 using Th17 cells cultured for 72h. Figures on the left show distribution of FOSL1 and FOSL2 binding sites relative to the position of the closest transcription start site (TSS). TSS is defined to be at position zero. The adjoining figure on the right is an overlay plot that compares the binding profiles of the two factors. ( C ) The topmost consensus sequences for FOSL1 and FOSL2 genomic-binding were identified using de-novo motif enrichment analysis by Homer. FOSL1 [left] and FOSL2 [right] peaks were further enriched for known TF motifs and the top six motifs identified by Homer are shown. Peaks with IDR p < 0.01 were used for motif discovery. ( D ) ChIPpeakAnno was used to determine the overlap in the genomic binding sites of FOSL1 and FOSL2 (overlap represents peaks sharing 200 bp or more). Genes neighboring to these overlying sites and differentially expressed under DKD or DOE conditions (FDR ≤ 0.1, |fold-change| ≥ 1.5) were assigned as the shared-direct targets of FOSL1 and FOSL2. Adjoining volcano plots show the logarithmic fold changes for selected shared targets (DKD [left] and DOE [right]). Downregulated genes are in blue, and upregulated ones are in red. Targets with FOSL occupancy over promoter regions (5-kb window around TSS) are highlighted in yellow. ( E ) Integrative Genomics Viewer (IGV) track snapshots show the binding overlap of FOSL1 and FOSL2 over selected Th17 genes.

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Immunofluorescence, ChIP-sequencing, Cell Culture, Binding Assay

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Bar plot depicts peak-annotation results for binding sites of FOSL1 and FOSL2 in 72h Th17-polarized cells. ( B ) Genes that were co-regulated (i.e., DE under DKD or DOE conditions) and showed co-localized genomic-binding of FOSL1 and FOSL2, were annotated as their shared direct targets. Venn diagram in the figure highlights (in bold) the shared targets [DKD, left; DOE, right] that are bound by FOSL factors over putative-promoter regions (5-kb around TSS). Out of these, the Th17-relevant targets have been marked in the volcano plots of .

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Binding Assay

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Heatmap on the top shows logarithmic fold-change values for the DE genes that are oppositely regulated in FOSL DKD and BATF KD Th17 cells, at the indicated time points of polarization. Heatmap in the bottom panel depicts the DE genes that are similarly altered in FOSL DOE and BATF KD Th17 cells. Th17-relevant genes are highlighted in red. ( B ) Comparing the ChIP-seq profiles of FOSL1, FOSL2 and BATF in Th17 cells. Graph (above) shows the overlay between the peak distribution profiles of the three TFs. Bar plot (below) depicts peak-annotation results for their identified binding sites. ( C ) Heatmap with k-means clustering shows the ChIP-seq signal intensities ± 2-kb around the centers of the genomic-binding regions of FOSL1, FOSL2 and BATF. Th17-associated genes in the vicinity of the binding sites are highlighted within the respective clusters. ( D ) Venn diagram shows an overlap between the genomic binding sites of FOSL1, FOSL2 and BATF (overlap represents peaks sharing 200 bp or more). Adjoining heatmap depicts fold-change values for the gene targets that are co-bound and oppositely regulated by FOSL proteins and BATF. Genes showing shared occupancy of the three factors over putative-promoters have been marked (*asterisk). Th17-relevant targets are highlighted. ( E ) IGV track snapshots illustrate the co-localization of FOSL1, FOSL2 and BATF over selected Th17 genes. Profile of H3K27ac marks around the shared sites is shown. ( F ) Bar plot depicts immunoblot-based expression analysis of STAT4 in FOSL DKD [left] and BATF KD [right] Th17 cells, cultured for 72h. Data shows mean ± SEM for three or four biological replicates, as indicated. Statistical significance is calculated using two-tailed Student’s t test (*p < 0.05). Adjoining IGV track shows the binding overlap of FOSL1, FOSL2 and BATF, flanked by H3K27ac marks near the STAT4 locus.

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: ChIP-sequencing, Binding Assay, Western Blot, Expressing, Cell Culture, Two Tailed Test

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Figure illustrates the common binding partners of FOSL1 and FOSL2 in Th17 cells (72h), based on data acquired from a parallel study of our lab ( bioRxiv ). Interactors having reported roles in T-cell function are shown. ( B ) STRING network analysis of human BATF. Width of lines between the nodes indicate confidence values for each protein-protein association. Only interactions with a minimum score of 0.7 are shown (high confidence). ( C and D ) Immunoprecipitated BATF was analyzed for its interaction with selected common binding partners of FOSL1 and FOSL2 (JUNB, SIRT-1, JUN and RUNX1), using western blotting (panel C). Additionally, BATF-interaction with STAT3 and IRF4 was analyzed to validate their previously-known association in mouse (panel D). Data is shown for three biological replicates.

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Binding Assay, Cell Function Assay, Immunoprecipitation, Western Blot

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Enrichment of disease-associated SNPs (or their proxies in Caucasian populations) within FOSL1, FOSL2 and BATF genomic-binding sites, relative to random sets of background SNPs. ( B ) SNPs relevant to the study were shortlisted. Out of these, the SNPs that were functionally validated in DNA-affinity precipitation assays (DAPA) have been shown. ( C and D ) DAPA reveals the SNPs that alter the binding of FOSL1, FOSL2 or BATF to their genomic sites that were identified by ChIP-seq analysis. Wildtype (WT) oligonucleotides containing the binding motifs of these TFs (at different genomic loci), and mutant oligonucleotides harboring a SNP within the corresponding motif, were used as baits. For experimental controls, an oligonucleotide with a conserved binding sequence for BATF (BATF WT), and the corresponding mutated sequence which is known to disrupt BATF occupancy (BATF MUT) were used. Immunoblot results for the SNPs unique to FOSL1, FOSL2 and BATF (panel C), and the ones common across the three factors (panel D) are shown. Data is representative of three biological replicates.

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Binding Assay, Affinity Precipitation, ChIP-sequencing, Mutagenesis, Sequencing, Western Blot

Journal: bioRxiv

Article Title: The AP-1 factors FOSL1 and FOSL2 co-regulate human Th17 responses

doi: 10.1101/2021.04.26.441472

Figure Lengend Snippet: ( A ) Table illustrates information on the autoimmune-linked SNPs that are harbored within consensus AP-1 motifs at the shared genomic-binding sites of FOSL1, FOSL2 and BATF. The sequence logos shown have been derived from the respective TF ChIP-seq peaks using Homer. ( B and C ) DAPA analysis was performed to test the effect of selected SNPs on the DNA-binding abilities of FOSL1, FOSL2 and BATF. The immunoblot images in panels B & C show biological replicates (R1, R2, R3) for and respectively

Article Snippet: To generate linearized vectors for the IVT reaction, the T7 promoter containing plasmids: empty pGEM-GFP64A, pCMV6-AC-GFP-FOSL1 (Origene, Cat no. RG202104) and pCMV6-AC-GFP-FOSL2 (Origene, Cat no. RG204146), were in vitro digested using the restriction enzymes Spe1 (NEB, Cat no. R0133), Xma1 (NEB, Cat no. R0180) and Ssp1 (NEB, Cat no. R3132), respectively.

Techniques: Binding Assay, Sequencing, Derivative Assay, ChIP-sequencing, Western Blot

Journal: Molecular Psychiatry

Article Title: AP-1 controls the p11-dependent antidepressant response

doi: 10.1038/s41380-020-0767-8

Figure Lengend Snippet: a A schematic diagram showing the timeline for biochemistry and behavior experiments during chronic SSRI fluoxetine (Flx) treatment. Fluoxetine was administered orally to BALB/cJ mice for 28 days. Start of treatment indicated as Day 1 (red arrow). Behavioral analysis (black arrows) using open-field test (OFT) was performed on day 14 (Supplementary Fig. ), tail suspension test (TST) on day 16, and novelty suppressed feeding test (NSF) on day 18. Biochemistry was done after harvesting the mouse prefrontal cortex (PFC) on 2, 5, 9, 14, 21, and 28 days of treatment (blue arrows). The onset and maintenance of behavioral response is shaded in gray. b TST. c NSF. d – k Quantitative-polymerase chain reaction (qPCR) to measure the levels of transcription factor mRNA Creb1 ( d ), Crem ( e ), Egr1 ( f ), c-Fos ( g ), Fosl2 ( h ), c-Jun ( i ), Junb ( j ), Jund ( k ). l A schematic diagram describing the formation of the AP-1 complex. Extracellular signals activate c-Fos and c-Jun mRNA transcription and protein expression. The proteins get phosphorylated by kinases, forming a stable heterodimeric AP-1 complex, thereby binding to target DNA and controlling their transcription. m A schematic diagram depicting c-Fos and c-Jun regulation by chronic fluoxetine treatment. Statistical analysis was performed between vehicle- (1% saacharine in drinking water) and fluoxetine-treated samples using two-tailed unpaired Student’s t -test; n = 6 for biochemistry and n = 11-14 for behavioral experiments. Data are mean ± SEM; * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.005.

Article Snippet: Creb1 , Mm00501607_m1, Rn00578826_m1; Crem , Mm04336053_g1, Rn04338541_m1; Egr1 , Mm00656724_m1, Rn00561138_m1; c-Fos , Mm00487425_m1, Rn00487426_g1; Fosl2 , Mm00484442_m1, Rn00564121_m1; Fosl1 , Mm04207958_m1, Rn00564121_m1; Fosb , Mm00500401_m1, Rn00500401_m1; c-Jun , Mm00495062_s1, Rn99999045_s1; Junb , Mm04243546_s1 Rn00572994_s1; Jund , Mm04208316_s1, Rn00824678_s1; S100a10 , Mm00501457_m1, Rn01409218_m1; Bdnf , Mm04230607_s1, Rn02531967_s1; Fgf2 , Mm00433287_m1, Rn00570809_m1; Egf , Mm00438696_m1, Rn00563336_m1; Igf , Mm00439560_m1, Rn00710306_m1; Ngf , Mm00443039_m1, Rn01533872_m1; Vegf , Mm00437306_m1, Rn01511602_m1; Tgfβ , Mm01178820_m1, Rn00572010_m1; Gapdh , Mm_99999915_g1, Rn99999916_s1; Srsf5 , Mm00833629_g1; Slc1a2 , Mm01275814_m1; Sirt1 , Mm01168521_m1; Glul , Mm00725701_s1; Glo1 , Mm00844954_s1; Crhr1 , Mm00432670_m1; Adrb1 , Mm00431701_s1; Abcb1 , Mm00440736_m1.

Techniques: Suspension, Real-time Polymerase Chain Reaction, Expressing, Binding Assay, Two Tailed Test

Journal: Molecular Psychiatry

Article Title: AP-1 controls the p11-dependent antidepressant response

doi: 10.1038/s41380-020-0767-8

Figure Lengend Snippet: Identification of transcription factor(s) regulating p11 by RNAi.

Article Snippet: Creb1 , Mm00501607_m1, Rn00578826_m1; Crem , Mm04336053_g1, Rn04338541_m1; Egr1 , Mm00656724_m1, Rn00561138_m1; c-Fos , Mm00487425_m1, Rn00487426_g1; Fosl2 , Mm00484442_m1, Rn00564121_m1; Fosl1 , Mm04207958_m1, Rn00564121_m1; Fosb , Mm00500401_m1, Rn00500401_m1; c-Jun , Mm00495062_s1, Rn99999045_s1; Junb , Mm04243546_s1 Rn00572994_s1; Jund , Mm04208316_s1, Rn00824678_s1; S100a10 , Mm00501457_m1, Rn01409218_m1; Bdnf , Mm04230607_s1, Rn02531967_s1; Fgf2 , Mm00433287_m1, Rn00570809_m1; Egf , Mm00438696_m1, Rn00563336_m1; Igf , Mm00439560_m1, Rn00710306_m1; Ngf , Mm00443039_m1, Rn01533872_m1; Vegf , Mm00437306_m1, Rn01511602_m1; Tgfβ , Mm01178820_m1, Rn00572010_m1; Gapdh , Mm_99999915_g1, Rn99999916_s1; Srsf5 , Mm00833629_g1; Slc1a2 , Mm01275814_m1; Sirt1 , Mm01168521_m1; Glul , Mm00725701_s1; Glo1 , Mm00844954_s1; Crhr1 , Mm00432670_m1; Adrb1 , Mm00431701_s1; Abcb1 , Mm00440736_m1.

Techniques: