Fig. 3

PNPLA2/ATGL is responsible for LDs mobilization upon BETi treatment. A, B Gene and protein expression in MDA-MB231 and Hs578t after 1 μM BETi treatment for the indicated time. A Left, RT-qPCR expression analysis on lipid metabolic genes selected in Fig. 1I after 24 hours of BETi treatment. Right, western blot analysis showed ATGL and DGAT1 protein expression in MDA-MB231. B Left, RT-qPCR results on PNPLA2 and DGAT1 genes in Hs578t after 24 hours of BETi treatment. Right, western blot analysis showed ATGL and DGAT1 protein expression in Hs578t. C Representative immunofluorescence images on MDA-MB231 and Hs578t treated with 1 μM of BETi for 24 hours. Green is BODIPY 500/510; red is ATGL, and blue is DAPI (nuclei staining). D Flow cytometry analysis of LDs content stained with BODIPY 500/510 in MDA-MB231 and Hs578t after 24 hours of BETi treatment at different concentrations (from 0.2 to 2 μM) and, below, ATGL expression analysis in both cell lines after 24 hours of BETi treatment from 0.1 to 2 μM. E Representative immunofluorescence images of MDA-MB231 and Hs578t cells after 6 days of treatment with 40 μM ATGListatin (green indicates LDs stained with BODIPY 500/510). F On top, a schematic timeline of the treatment. Below, a flow cytometry analysis of LDs content stained with BODIPY 500/510 in MDA-MB231 and Hs578t cells treated with ATGListatin and BETi. bACT is used as a loading control in both cell lines