Article Title: Arid1a loss potentiates pancreatic β-cell regeneration through activation of EGF signaling
Figure Lengend Snippet: Arid1a deficient β-cells have a dependence on increased EGF/Neuregulin signaling. A. Heatmap of differentially expressed genes in WT and KO islets from Arid1a Fl/Fl and Ubc-CreER; Arid1a Fl/Fl mice, isolated after PPx and detected by RNA-Seq (left). Heatmap showing a subset of overexpressed genes in KO islets (right; n = 2 and 2 mice). B. GSEA shows that EGF and NRG1 response genes are upregulated in KO islets. The nominal enrichment score (NES), nominal p-value, and false discovery rate (FDR) q-value are shown within each GSEA plot. C. p-EGFR western blots in MIN6 and BTC6 treated with different doses of EGF. D. p-EGFR western blots in WT MIN6 cells treated with EGF in the presence or absence of erlotinib and canertinib. E. Relative cell numbers for control shGFP and shArid1a MIN6 cells in the presence of canertinib and erlotinib. Shown as % of control cells ( shGFP group treated with DMSO). F. Western blot showing ARID1A levels in MIN6 clones. MIN6 cells were transduced with lenti- CAS9 -blasticidin and either non-targeting lenti-sgRNA ( Gal4 )-puromycin or lenti-sgRNA ( Arid1a )-puromycin. G. MIN6 clone growth over 15 days measured by cell counting. H. mRNA expression of selected genes in Gal4 control and Arid1a KO MIN6 clones as measured by qPCR. I. K-mean clustering of H3K27Ac ChIP-Seq peaks in non-targeting Gal4 , Arid1a heterozygous and Arid1a KO MIN6 clones. J. Annotation of clusters defined by H3K27Ac ChIP-Seq sites by distance to TSS. K. Sample tracks for H3K27Ac ChIP-Seq in control, Arid1a heterozygous and Arid1a KO MIN6 single cell clones at the Nrg4 and Egfr loci.
Article Snippet: Either 20 mg/kg canertinib or 20 mg/kg NR4A1 antagonist C-DIM8-DIM-C-pPhOH (Axon Medchem # Axon 2827) was administered daily through gavage starting a day before PPx until sacrificing mice day 7 post-PPx.
Techniques: Mouse Assay, Isolation, RNA Sequencing Assay, Western Blot, Clone Assay, Transduction, Cell Counting, Expressing, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation