Fig. 5

Blocking the Shh pathway can reverse the stimulatory effect of SCUBE1.(A) Coimmunoprecipitation of SCUBE1 and Shh in PLC and Huh7 cells. The IgG isotype was used as a negative control. (B) The invasion capacity of PLC and Huh7 cells treated with rhSCUBE1 and IgG was detected with Transwell assays (**P < 0.01, t test). Scale bar, 200μm. (C) The expression of CD133 in different groups was measured via Western blotting. (D) The Shh content in the supernatant in PLC and Huh7 cells with different treatments was measured with ELISA (*P < 0.05, t test). (E) The proliferation rates of PLC and Huh7 cells after the addition of rhSCUBE1 with or without Shh antibody were measured (***P < 0.001 vs. rhSCUBE1, one-way ANOVA). (F) Western blot analysis of Smo and Gli1 expression in PLC and Huh7 cells treated with rhSCUBE1 and cyclopamine. β-Actin was used as an internal control. (G) The invasion capacity of PLC and Huh7 cells treated with rhSCUBE1 and cyclopamine was measured with Transwell assays (**P < 0.01, t test). Scale bar, 200μm. Data are represented as mean ± SD of three independent experiments