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. 2015 May 9;8(48):83457–83468. doi: 10.18632/oncotarget.4054

Figure 2. SH-I-14 disrupts the STAT3-DNMT1 interaction.

Figure 2

A. SH-I-14 inhibits the STAT3-DNMT1 interaction in HS578T and SUM1315MO2. Cells were treated with increasing amount of SH-I-14 for 24 hr and STAT3-bound complexes were precipitated by anti-STAT3 antibody. Western blot analysis was performed with indicated antibodies. B. JAK1/2 inhibitors do not disrupt STAT3-DNMT1 interaction. HS578T cells were treated with increasing amount of compounds for 24 hr and immunoprecipitation/western blot analysis was performed as in A.. C. STAT3-DNMT1 interaction is dependent on the acetylation of STAT3 at K685. GFP-tagged wild type STAT3, STAT3 (K685Q; KQ), or STAT3 (K685R; KR) was transfected into PC-3 cells. After immunoprecipitation by anti-STAT3 antibody, western blot analysis was performed with indicated antibodies. D. SH-I-14 disrupts wild type STAT3-DNMT1 interaction but not STAT3 KQ-DNMT1 interaction. PC-3 cells were transfected with either GFP-STAT3 WT or GFP-STAT3 KQ and STAT3 proteins were immunoprecipitated by anti-STAT3 antibody. β-actin was used as a loading control. Abbreviations: WT, wild type STAT3; KQ, STAT3 (K685Q); KR, STAT3 (K685R); WCL, whole cell lysates; IP, immunoprecipitation; and WB, western blot.