Fig. 6
BETi cooperate with FoxO1 to mediate ATGL transcriptional activation. A The top 5 GO (gene ontology) categories as in Fig. 1G and the plot of the log2 fold change value of FoxO signaling pathway category; red indicates FoxO1. B Time-course (30 minutes to 24 hours) protein expression analysis of FoxO1 and ATGL in MDA-MB231 and Hs578t after 1 μM JQ1 treatment. C RT-qPCR analysis of FoxO1 and ATGL (PNPLA2 gene) after 1 μM JQ1 treatment at different time points (30 minutes to 24 hours). D ChIP analysis of FoxO1 binding on PNPLA2 promoter after 6 hours of 1 μM BETi treatment in MDA-MB231, Hs578t and MCF7 cell lines. The TAAACT FoxO1 binding site is located around 1.6Kb downstream of the TSS. The line indicates the IgG value E Protein expression analysis of ATGL and FoxO1 in control (siCNT) and FoxO1 siRNA cells (siFoxO) after 48 hours of siRNA transfection combined with 1 μM BETi for the last 24 hours. F Western blot analysis of FoxO1 and ATGL in MDA-MB231 and Hs578t in control (EV) and FoxO1 overexpression (FoxO) after 24 hours from transfection. Representative images are selected for the western blot. bACT served as a loading control