Extended Data Figure 6. Autophagy regulates primary ciliogenesis in a cell cycle independent manner.
a, FACS analysis of Atg5+/+ and Atg5−/− MEFs in normal medium or subjected to 24-hour serum starvation. Data shown represent for 1×106 cells per well in triplicate samples. b, Primary ciliogenesis is less efficient when the lysosome activity is blocked in MEFs. Representative confocal images of primary cilia formed in Atg5+/+ MEFs subjected to 24-hour serum starvation alone or combined with 20 μM CQ treatment. c, Quantified percentage of cells with primary cilia in b. d, Quantified length of primary cilia in b. e, Degradation of Ofd1 is also blocked in Atg3 −/− MEFs. Western blot analysis of Ofd1, p62, LC3-I/II and BBS4 protein levels in MEFs with indicated genotypes in normal medium or subjected to 24-hour serum starvation, quantified Ofd1 levels were normalized with β-tubulin. f, g, Primary ciliogenesis is also defective in Atg3−/− MEFs. f, Quantified percentage of cells with primary cilia in Atg3+/+ and Atg3−/− MEFs in normal medium or subjected to 24-hour serum starvation. g, Quantified length of primary cilia formed in Atg3+/+ and Atg3−/− MEFs as described in f. c, d, f, g, Data shown represent mean ± s.d. for 100 cells per well in triplicate samples. ***P<0.001, two-tailed unpaired student’s t-test. a–g, Similar results were obtained in three independent experiments.