Fig. 3: SMARCB1 loss upregulates STAT3 expression.

A, B Gene set enrichment analysis (GSEA) of RNA-seq data from orthotopic tumors derived from T24 control, SMARCB1 KO and SMARCB1 rescue xenografts. Volcano plot represents the hallmark pathways that were enriched in A SMARCB1 KO over T24 control (FDR < 0.25) and B SMARCB1 rescue over SMARCB1 KO (FDR < 0.25). C Relative fold change in mRNA levels of STAT3 from T24 ctrl (n = 6 replicates sampled across three different mice; each one analyzed under three technical replicates), SMARCB1 KO (n = 6 replicates sampled across three different mice; each one analyzed under three technical replicates) and SMARCB1 rescue (n = 6 replicates sampled across three different mice; each one analyzed under three technical replicates) orthotopic xenografts. Normalized with β-actin [Data are represented as mean ± standard deviation (SD)]. D Immunoblot analysis of pSTAT3 (Y705), STAT3, SMARCB1 in T24 ctrl, SMARCB1 KO and SMARCB1 rescue orthotopic tumors (n = 3). Same lysate was used for upper and bottom panels. β-actin was used as loading control. Scatter plots show relative expression of the target proteins after background subtraction and normalization to β-actin [Data are represented as mean ± standard deviation (SD)]. E ChIP-qPCR analysis shows increased levels of H3K4me3 and H3K27ac on STAT3 promoter in SMARCB1 KO xenografts (n = 4; two biological replicates analyzed under two technical replicates) [Data are represented as mean ± standard deviation (SD)]. F ChIP-qPCR analysis shows decreased levels of SMARCB1 on STAT3 promoter in SMARCB1 KO xenografts which was rescued in SMARCB1 re-expression (n = 6; three biological replicates analyzed under two technical replicates) [Data are represented as mean ± standard deviation (SD)]. G Immunoblot analysis of pSTAT3 (Y705), STAT3, and SMARCB1 in T24 ctrl, KO, and rescue spheroids treated with the JAK1 inhibitor, Itacitinib for 10 days at 1000 nM concentration. β-actin was used as loading control. For panels C, D, E and F, P-values were determined by unpaired two-tailed Student’s t-test. Source data are provided as a Source Data file.