Fig. 5.
T2D plasma exosomes require BRD4 to upregulate EMT genes and PD-L1 expression. (A) Expression of SNAI1 and CD274 genes in DU145 cells measured by RT-PCR of cellular RNA after exosome treatment. T2D exosomes (109; T2D Exo) were compared to media-only control exosomes (Control), or T2D Exo + JQ1 or MZ1. (JQ1 is a pan-BET inhibitor (400 nM) and MZ-1 is a BRD4-selective degrader (50 nM)). (B) Expression of PD-L1 was measured by flow cytometry after treatments. One million events were analyzed by Flow-Jo and presented as overlaid histograms (control, red trace; experimental, black trace). (C) Flow cytometry data of B were quantified with PD-L1 as percent of parent population. The experiment was conducted in triplicate with differences between means represented as bar graphs. Data were analyzed by two-way ANOVA with statistical significance presented as: ****, P < 0.0001 (TGFB, TGF-β positive control at 5 ng/mL for SNAI1 induction; IFNgamma, interferon-γ positive control at 5 ng/mL for CD274 induction). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)