Fig. 5

ATGL downregulation influences mitochondrial and metabolic dynamics in TNBC cells. A Western blot analysis showing ATGL downregulation in MDA-MB231 and Hs578t in control (siCNT) and ATGL siRNA cells (siATGL) after 48 and 72 hours from siRNA transfection. B DHE fluorescence measure to evaluate the ROS content after 48 hours of ATGL downregulation. C, D Protein expression analysis of SOD1 and ATGL (C) and (D) SOD1 band quantification in MDA-MB231 and Hs578t cells after 48 h of ATGL downregulation. E-F Protein expression analysis of CPT1a and ATGL (E) and (F) CPT1a band quantification in MDA-MB231 and Hs578t cells after 48 h of ATGL downregulation. G RT-qPCR analysis to evaluate the effect of ATGL downregulation (siRNA 48 h) on glycolytic (upper panel) and β-oxidation (bottom panel) genes in MDA-MB231 and Hs578t. H MTT assay to evaluate the mitochondrial metabolic activity in MDA-MB231 and Hs578t after 48 h of siATGL transfection. I-L Schedule: 48 h of siRNA transfection (siCNT and siATGL) and the last 24 h 1 μM of BETi treatment in MDA-MB231 and Hs578t. I ATGL expression analysis and (J) flow cytometry analysis of LDs content stained with BODIPY 500/510. K MTT assay with two different doses (0.1 and 1 μM) of BETi. L CPT1a protein expression analysis. Representative images are selected for the western blot. bACT served as a loading control