LC3C inhibits asymmetric inheritance of PDMBs. (A) Examples of asymmetric cell division in different phases of cell division in 786-O VHL(+) and UOK 257 FLCN(+) LC3C-knockdown cells. Arrowheads point to the PDMBs, which are different from the cytokinetic midbody in the intercellular bridge. Scale bars = 10 µm. (B) Percentage of symmetric and asymmetric cell divisions compared with the total number of cell divisions in 786-O VHL(+) cells without and with reconstituted VHL. (C) Percentage of symmetric and asymmetric cell divisions compared with the total number of cell divisions in Caki-1 cells with endogenous VHL and with VHL knocked down. (D) Percentage of symmetric and asymmetric cell divisions compared with the total number of cell divisions in UOK 257 cells without and with FLCN. (E) Knockdown of LC3C increased the percentage of asymmetric versus symmetric cell divisions compared with the total number of cell divisions in Caki-1 cells with LC3C knocked down. (F) 786-O VHL(+) cells with LC3C knockdown grown as three-dimensional spheroids maintained higher levels of PDMBs (CYK staining) in multiple daughter cells. (G) Quantification of PDMBs, nuclei, and ratios of PDMBs/nuclei for 20 spheres in each category. Scale bars = 20 µm. (H) Immunocytochemistry for PDMBs (CYK4 staining; arrowheads point to PDMBs) and quantification of PDMBs in sections from human normal kidney cells and human ccRCC (hccRCC) cells with WT or mutated (mut) VHL. Magnified inset scale bars = 10 µm. In all bar graphs, mean ± SEM is shown. *, P < 0.05; **, P < 0.01; ***, P < 0.001 by unpaired two-tailed t test. A, asymmetric; KD, knockdown; S, symmetric; Scr, scramble.