Fig 1: USP22 mediates PIC stability at ER stress response genes ASchematic of the CHOP locus. Black boxes indicate amplicons for subsequent ChIP-qPCR analysis.BHCT116 cells were treated for 2 h with 100 nM Thapsigargin in the presence or absence of shRNA targeting USP22. Cells were cross-linked, harvested, and subjected to ChIP-qPCR for total RNA Pol II, pSer5 Pol II, and pSer2 Pol II at the loci across CHOP (indicated in (A)). Three independent experiments are represented as mean ± SEM.CChIP-qPCR for total RNA Pol II, pSer5 Pol II, and pSer2 Pol II at ERP70, GRP78, and a gene desert (Prom = gene promoter and Coding = downstream coding region—see Appendix Table S1 for primer sequences). Three independent experiments are represented as mean ± SEM. with significance measured by Student’s t-test. *P < 0.05, **P < 0.01, and ***P < 0.005.D–GCells were treated as in (B) and subjected to ChIP-qPCR for TBP, TFIIB, TFIIF, and TAF7 at the indicated gene promoters. Three independent experiments are represented as mean ± SEM, with significance measured by Student’s t-test. *P < 0.05 and ***P < 0.005.HAn aliquot of cells from (D-G) were harvested for whole-cell lysate and subjected to immunoblotting with the indicated antibodies. Source data are available online for this figure.