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. 2021 Jun 8;11:642134. doi: 10.3389/fonc.2021.642134

Figure 3.

Figure 3

BRD4 regulates the activation of the JAK/STAT3 signaling pathway through GP130 receptor. (A) NHWD-870 directly inhibits the phosphorylation of STAT3 at tyrosine 705 in HOS and 143B osteosarcoma cell lines. (B) Under IL-6 stimulation, the STAT3 signaling pathway was significantly activated, and NHWD-870 effectively inhibited IL-6-induced STAT3 phosphorylation activation. (C) The expression levels of p-JAK and p-STAT3 were quantified using ImageJ software. (****P < 0.0001). (D) The nuclear translocation of STAT3 in 143B cells was determined using immunofluorescence. Scale bar = 50 μm. (E) The gene track for H3K27 acetylation and BRD4 in HepG2 and k562 cells from ENCODE. (F) ChIP-PCR for BRD4 association with the TSS of GP130 in 143B osteosarcoma cells treated with 1 µM NHWD-870 or DMSO using anti-BRD4 or anti-IgG as negative control. Data are expressed as the fold enrichment normalized to the input. (****P < 0.0001).