Smo activates autophagy via AMPK, and promotes Gα_i activation to induce ciliogenesis. (A) Western blots indicating the efficiency of different KDs in RPE1 cells as indicated. OFD1 siRNAs represent a combination of different OFD1 siRNAs as indicated. (B) Quantification of ciliated cells for depletion of different Gα_i isoforms using siRNA in RPE1 cells. (C) Immunofluorescence images of RPE1 cells plated at LCD and treated with Rp-8-Bromo cAMPs in the presence and absence of Shh. (D) Western blot probing for Akt total and phosphorylated to indicate the presence or absence of active mTOR upon Hh activation, respectively. (E) Western blot probing for total and phosphorylated acetyl-CoA carboxylase (ACC) to indicate the activation of AMPK upon Hh activation. (F) Immunofluorescence images of RPE1 cells plated at LCD and treated with SAG or SAG and Torin1 for 24 h.*, P < 0.01; **, P < 0.001, ***, P < 0.0001, comparing all conditions to control cells with Shh. Number of cells (n) used for each experiment (three repeats) is listed in Table S4. Yellow and red arrows indicate the presence or absence of a cilium, respectively.