Fig. 2

APOL1 repression controls tumor progression and lipid deposition in VHL^(−/−) ccRCC cells. (A, B) VHL^(−/−) ccRCC (786-O, A498 and OS-RC-2) were transfected with two independent shRNAs against APOL1 or a control (Lacz). qRT-PCR and Western blot analysis of APOL1 are shown. (C) GSEA assays for the correlation of fatty acid metabolism, adipogenesis and mRNA levels of APOL1 according to the TCGA database. FDR < 25%, p < 0.05 was considered statistically significant. (D) APOL1 protein levels were detected by immunoblot analysis in renal cancer cell lines. GAPDH served as an internal control. (E) The correlation between APOL1 protein and the quantification of ORO in renal cancer cell lines and HK-2 cell line. (F) Photomicrographs of Oil Red O-stained VHL^(−/−) cells 786-O, A498 and OS-RC-2 with APOL1 knockdown (Magnification: 200× & 400×). (G, H) Quantification of ORO, relative diameter of lipid droplets in indicated cell lines. The data are presented as the means ± SEM. p values of two-tailed Student’s t tests are displayed. (I) Representative micrographs of crystal violet-stained cell colonies analyzed by clonogenic formation. (J) MTS assays revealed cell growth curves of indicated cells. (K) Migration and invasion assays for indicated renal cancer cells. Representative photographs were taken at 200× magnifcation; the number of migrated cells was quantified in three random images from each treatment group