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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Clin Cancer Res. 2019 Mar 13;25(13):4179–4193. doi: 10.1158/1078-0432.CCR-18-3535

Figure 3. Drugs affecting ciliation influence early events in SHH and PDGFRα signaling response.

Figure 3.

A. Steps in SHH signaling pathway activation. B., C. Representative images (B) and quantitation (C) of Smo entry into the cilia of NIH3T3 cells treated with indicated drugs for 1 hour, then exposed to medium +/− Shh-N (2 ng/ml), (n≥500 cells). Scale bar 5 μm. Green, Arl13b; red, Smo; blue, DAPI; images of Smo and Arl13b are offset. D., E. Representative images (D) and quantitation (E) of Gli3 distribution within the cilia of NIH3T3 cells treated with indicated drugs for 1 hour then exposed to medium +/− Shh-N (2ng/ml), (n≥500 cells). Top and middle panels: Green, Gli3; purple, Arl13b; blue, DAPI; images of Gli3 and Arl13b are offset, scale bar 5 μm. Bottom panel: γ-tubulin (magenta) indicates basal bodies and demonstrates localization of Gli3 (red) at the ciliary tip. Green, Arl13b, red, Gli3, magenta, γ-tubulin; blue, DAPI; images of Gli3 and Arl13b are offset, scale bar 2 μm. F., G. Representative Western blot images and quantitation of PDGF-AA-induced activation of PDGFR and MEK1/2 in NIH3T3 cells treated with indicated drugs for 2 h (F) or 30 min (G); phosphorylation is expressed based on normalization to total protein, (n=3, n, number of biological replicates). See also Suppl. Fig S4, for data normalized to loading control only. Significance calculated by Tukey test in all cases, from three independent experiments; *, p <0.5; **, p <0.01; ***, p<0.001, relative to vehicle.