Hh activation induces ciliogenesis in RPE1 cells and primary neurons. (A) Schematic indicating canonical and noncanonical Hh pathways. (B) Immunofluorescence images of RPE1 Ptch1:GFP cells treated with Shh. Ptch1 localized to cilia when Hh pathway was inactive and was removed from cilia upon Hh activation. (C) Immunofluorescence stitched images of RPE1 cells plated at different cell densities indicating a bigger field of view for comparison between different cell densities. (D) Immunofluorescence images of neurons from mouse cortex indicating staining with adenylate cyclase III (AC3) as another ciliary marker. (E) Immunofluorescence images of neurons and “nonneuron” cells in the same culture indicating lack or presence of YAP1 expression in neurons and nonneurons, respectively. Images are stitched to provide a bigger field of view. Yellow and red numbers and arrows indicate the presence or absence of a cilium, respectively.