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. 2016 Mar 2;6:22540. doi: 10.1038/srep22540

Figure 5. Compound 6 and 9 effect on PDAC tumor spheroids growth by antagonizing ciliary SmoM2-mediated Hh pathway activation.

Figure 5

(A) Illustration of experimental scheme to generate 3D tumor spheroids and for compounds administration. (B) Representative CLSM images and (3D) 3D reconstruction of whole-mount staining of spheroids after treatment with compound 6 or 9. PC are visualized with an antibody against Ace-Tub, and SmoM2 and Gprasp2 are visualized with antibodies to GFP and Gprasp2 respectively. Quantification of (C) size, (D) ciliation, and (E) ciliary SmoM2 and Gprasp2 of spheroids treated with compound 6 or 9. (F) Gli1 protein expression levels in spheroids treated with compound 6 or 9. Graphs indicate quantification of immunoblot data that show the mean fold change of protein normalized to α-Tub levels. (G) Statistical analysis of cell cycle proliferation of spheroids determined by BrdU incorporation assay after compound 6 or 9 treatment. Scale bar = 10 μm. >100 cilia and tumor spheroids were analyzed per condition. All error bars represent the mean SD of three independent experiments. A two tailed unpaired t-test is used for statistical data analysis. (*p < 0.01, #p < 0.001).