Figure 4.

PPARA requires autophagy for ligand-induced ciliogenesis. (A) MEFs with indicated genotypes were treated with 0.5 μΜ GW 7647 for 24 h, cells were immunostained with anti-ARL13B Alexa Fluor 568-conjugated antibody. The cells were observed using fluorescence microscopy. Data shown represent mean ± s. d. percentage of cells with primary cilia, for 200 cells per well in triplicate samples. (B) MEFs with indicated genotypes were treated with 0.5 μM GW 7647 for 24 hrs or 5 μM CQ for CQ for 12 h as indicated. After incubation, cells were lysed and immunoblotted with antibodies against LC3, SQSTM1, and ACTB. (C) MEFs with indicated genotypes were treated with 1 μΜ rapamycin for 24 h, cells were immunostained with anti-ARL13B Alexa Fluor 568-conjugated antibody. Data shown represent mean ± s. d. percentage of cells with primary cilia, for 200 cells per well in triplicate samples. (D) MEFs with indicated genotypes were treated with 1 μM Rapamycin as indicated. After 24 h, cells were lysed and immunoblotted with antibodies against SQSTM1, and ACTB. All the experiments were performed 3 times. *P < 0.05, **P < 0.01 and ***P < 0.001, one-way ANOVA, compared to that of cells incubated in normal, serum-free media or GW 7647 only.