Fig. 4

Influence of fatty acids (FAs) on cell proliferation. A Immunoblotting for ATGL and p21 in MDA-MB231 and Hs578t cells after treatment with 200uM of Oleic Acid (OA) and 5 mM of Propionic Acid (PA) with the respective vehicle (EtOH and H2O respectively) for 24 hours. B Cell cycle analysis of MDA-MB231 and Hs578t cells treated with OA and PA for 24 hours after 1 hour pulse of BrdU. BrdU and PI incorporation indicated the % of the population. Histograms are the % of cell population in all three different phases of the cell cycle. The significance is calculated versus the CNT treated cells. C, D Cell proliferation assay measured by IncuCyte to follow the effect of PA 5 mM (C) and OA 200uM (D) in MDA-MB231 and Hs578t in combination with 1 μM BETi treatment. E Left, immunoblotting for p21 in MDA-MB231 and Hs578t cells after ATGListatin treatment for 3 days. Right, cell cycle analysis of MDA-MB231 and Hs578t cells treated with ATGListatin for 3 days followed 1 hour pulse of BrdU. Histograms are the % of cell population in all three different phases of the cell cycle. F Left, cell proliferation assay measured by IncuCyte and, right, endpoint (6 days) to evaluate the effect of different ATGListatin concentrations in MDA-MB231 and Hs578t. G MTT assay to assess ATGListatin on mitochondrial metabolic activity after 6d of treatment at 40 μM. H Left, representative images of colony formation assay (cristal violet dye) in MDA-MB231 and Hs578t cells treated with ATGListatin 40 μM and right, quantification of the colonies by ImageJ sofware. bACT served as a loading control