Fig. 1

a–f MEF cells with or without serum starvation (SS) were treated with 5 mM 3-MA, or 20 μM CQ for 24 h, and applied for the following analyses. Western analysis showed the protein levels of endogenous CP110, LC3 and p62 (a). β-actin was used as a loading control. Immunofluorescence analyses were carried out by using anti-CEP164 and anti-CP110 antibodies (b). CEP164, a marker of mother centrioles. Scale bar, 2 µm. The percentage of cells with CP110 dot at mother centrioles was calculated (c). Immunostaining of γ-tubulin and ADP ribosylation factor like GTPase 13b (Arl13b) in the indicated cells is shown (d). Arl13b and γ-tubulin are cilia and centrosome markers, respectively. The arrowheads indicate cilia. Scale bar, 10 µm. The cells with cilia were counted (e). Cilia length was also measured using ImageJ software (f). g–l Autophagy-deficient Atg5^–/– or Atg7^–/– MEF cells cultured with or without serum were subjected to the following analyses. Western blotting showed the levels of the indicated proteins (g). Immunofluorescence analyses were carried out by using anti-CEP164 and anti-CP110 antibodies (h). Scale bar, 2 µm. The percentage of cells with CP110 dot at mother centrioles was counted (i). Immunostaining of γ-tubulin and Arl13b in the indicated cells is shown (j). The arrowheads indicate cilia. Scale bar, 10 µm. The ciliated cells were calculated (k) and cilia length was determined using ImageJ software (l). Quantitative data are expressed as the means ± SD (at least three independent experiments). n, sample size. *P < 0.05, **P < 0.01, and ***P < 0.001; ns, not significant (P > 0.05); Student’s t-test.