Fig. 6

STAiR18-mediated regulation of STAT3. a STAT3 mRNA and protein levels depend on STAiR18. KDs of STAiR18 and STAT3 were conducted in INA-6 cells, followed by RNA- and protein-isolation 24 h and 48 h, respectively. RNA expression was carried out by qPCR (n = 4); protein expression by SDS-PAGE and immunoblotting using a densiometric analysis (n = 4). The indication of significances refers to the control. The corresponding Western Blots are displayed in Additional file 1: Figure S9. b SETD2 is associated with STAiR18 and STAT3 mRNA. INA-6 cells were crosslinked and a RIP was performed using a specific SETD2 antibody together with an IgG negative control. RNA was analyzed by qPCR. Values were normalized to IgG control (n = 3). c Active chromatin state of the STAT3-locus depends on STAiR18. STAiR18 KD (targeting exon2) followed by ChIP was performed in INA-6 cells using antibodies targeting H3K4me3, H3K27me3, or H3K36me3, and SETD2 24 h after transfection. IgG was used as a negative control. Enriched DNA was isolated and analyzed by qPCR with specific primers for the STAT3 promoter region and the STAT3 gene. Values were normalized to the corresponding IgG negative control (n = 4)