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. 2012 Jul 10;109(30):12058–12063. doi: 10.1073/pnas.1206458109

Fig. 4.

Fig. 4.

The Sin3a complex negatively regulates STAT3 nuclear distribution and binding to the SOCS3 gene promoter. (A) Hek293T cells were transfected with control (Renilla luciferase) or SIN3A siRNA. After 72 h, cells were cultured 4 h without FCS and then stimulated with LIF for the indicated times. Cells were fixed and stained and the localization of STAT3 and Sin3a was assessed by confocal analysis. Immunofluorescence of representative cell fields is shown. (Magnification: 63×.) (B–E) Hek293T cells were stably transfected with a scrambled or a SIN3A-targeting shRNAmir. Transfection and silencing efficiency are reported in Fig. S3. Transfected cells were cultured 4 h without FCS and then stimulated with LIF for the indicated time points. ChIP assays were performed to examine the occupancy of Sin3a (B), STAT3 (C), phospho-pol II (D), and the acetylation status of histone 3 (H3) K27 acetylation (E) on the SOCS3 promoters. Immunoprecipitated DNA was quantified by quantitative PCR. Graphs represent occupancy levels relative to NS sample (fold-induction). Results are representative of three independent experiments. Error bars indicate SD from duplicates.