Fig 2: PB2 attenuated increase in Shh signalling pathway induced by CS. A, IHC staining and quantitative analysis of Shh, Smo and Gli1 in liver tissues, and scale bar represents 20 µm. B-D, Quantitative analysis of Shh, Smo and Gli1 in liver tissues. E, Expression of Shh, Smo and Gli1 protein in liver tissues. F-H, Quantitative analysis of Western blotting results. Similar results were obtained from three independent experiments. All data are presented as the mean ± SEM (n = 6 in each group). *P < 0.05 and **P < 0.01 vs Control group; ## P < 0.01 vs CS group

Fig 3: Rab1A upregulated Gli1 expression in an SMO-independent manner. (A–C). Gli1 and SMO expression in Rab1A-silenced cells as detected by Western blotting (A). The bands were quantified and presented as the mean ± S.E.M of three independent experiments (B and C). (D–F). Gli1 and SMO expression in Rab1A-overexpressed cells as detected by Western blotting (D). The bands were quantified and presented as the mean ± S.E.M of three independent experiments (E and F). (G) Gli1 expression is detected by Western blotting in Rab1A-overexpressed SW480 cells treated with cyclopamine (Cyc; 5 µM) for 24 h. Here cropped blots were displayed and all full-length blots are included in the Supplementary Information file. ns, nonsignificant, **P < 0.01.

Fig 4: Rab1A promoted cell proliferation and migration through a Gli1-dependent pathway in SW480 cells. (A) Gli1 expression is detected by Western blotting in Rab1A-overexpressed SW480 cells treated control siRNA (siControl) or Gli1 siRNA (siGli1). (B) Cell proliferation ability was detected by CCK8 assays in Rab1A-overexpressed SW480 cells treated control siControl or siGli1. (C) Migration ability was detected by the Transwell assays in Rab1A-overexpressed SW480 cells treated control siControl or siGli1. Here cropped blots were displayed and all full-length blots are included in the Supplementary Information file. ns, nonsignificant, **P < 0.01, ***P < 0.001.

Fig 5: ETV4 loss retarded growth of breast tumor xenograft.A Efficient transduction of MDA-MB-231 cells by GFP-expressing lentiviral vector carrying short shRNAs for ETV4 as reflected by green fluorescence. B, C Establishment of stable ETV4 knockdown MDA-MB-231 cells as confirmed by quantitative RT-PCR (B) and western blotting assay (C). D–F Nude mice (n = 6 each group) were subcutaneously implanted with control or shETV4 MDA-MB-231 cells. Tumor volumes were measured (D). Twenty-five days after inoculation, tumors were isolated (E) and weighted (F). G Immunoblots of ETV4, CXCR4, SHH and GLI1 in harvested MDA-MB-231 tumors. Data are presented as mean ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001.