Fig. 6

APOL1-dependent lipid storage is required for ER homeostasis and cell viability in ccRCC. (A) Heatmap showing the expression change of genes in 786-O cells after transfection of APOL1 shRNA and control shRNA Lacz. Gene expression is shown as RPKM after normalization. (B) GO enrichment for the indicated cells based on the results from sequencing. (C) KEGG enrichment top 20 for indicated cells based on the results from sequencing. (D, E) Q-RT-PCR analysis of UPR target genes in renal cancer cell 786-O and A498 with APOL1 knockdown. (F) 786-O and A498 cells were double immunostained with anti-APOL1 antibody (green) and ER-Tracker probe (red). The cell nuclei were counterstained with DAPI (blue). The co-localization between the protein and endoplasmic reticulum is shown in the merge panel (Magnification: 640×). (G) Western blot for UPR sensors was performed in 786-O and A498 cells with APOL1 knockdown. (H, I) ER-Tracker Red (500 nmol/L) staining of live cells described was performed. Representative images (left) and quantification of ER Tracker fluorescence are shown (right). Fluorescence was normalized to forward scatter for each event to account for differences in cell size (Magnification: 640×). P values were determined by the Students’ t test. (J) Transmission electron microscopy (TEM) of control and APOL1-depleted cells is shown. Red arrows, rough ER. (Magnification: 5000×)