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. 2021 Nov 15;32(2):190–209. doi: 10.1038/s41422-021-00584-9

Fig. 8. Enhanced ciliation in C11ORF49/CSTPP1- and TPGC- deficient cells results from defects in an actin-mediated cilium disassembly pathway.

Fig. 8

a Control and C11ORF49/CSTPP1−/− RPE-1 cells were serum-starved for 48 h and then re-stimulated with 10% FBS for the indicated times. Ciliation ratio and cilium length were measured and quantified at the indicated times. b Cycling control and C11ORF49/CSTPP1−/− RPE1 cells were infected with empty (EV), GFP-CENTRIN1, or GFP-CENTRIN1-VCA lentiviruses for 72 h and immuno-stained with indicated antibodies to examine ciliation and nuclear shape. c, d. Cycling control and C11ORF49/CSTPP1−/− RPE1 cells were infected with empty (EV), or Flag-MAP4m-VCA lentiviruses for 72 h and immuno-stained with indicated antibodies to examine ciliation and actin organization. e Cycling control and C11ORF49/CSTPP1−/− RPE1 cells were immuno-stained with indicated antibodies to examine YAP/TAZ localization. f Protein levels for PLK1 were examined in lysates from controls and cells depleted of indicated proteins by western blotting using indicated antibodies. g Cycling control and C11ORF49/CSTPP1−/− RPE1 cells were infected with or without GFP-YAP5SA for 72 h and immuno-stained with indicated antibodies to examine ciliation. Arrows indicate ciliated C11ORF49/CSTPP1−/− cells without expression of GFP-YAP5SA. h The intensity of centrosomal NDE1 was quantified in cycling, ciliated control RPE1 cells and cells depleted of TTLL1 or C11ORF49/CSTPP1 by IF using indicated antibodies. Scale bars, 2 μm in a and h. Scale bar, 10 μm in all other panels. n ≥ 60 per sample were analyzed in three independent experiments in a, f. n ≥ 100 per sample were analyzed in three independent experiments for all other data in this figure. All data are presented as means ± SD. *P < 0.05 (unpaired t-test or Mann-Whitney test).