Fig. 4

Activation of RAP1/CDC42 signaling is required for TTC17 deficiency to facilitate metastasis. a Profile of DEGs (logFC > 1.5, FDR < 0.05) between TTC17-knockout MDA-MB-231 cells and control cells. b Enrichment of GO biological processes for up- and downregulated genes. c Enrichment of KEGG pathways for up- and downregulated genes. d Heatmap of key molecules in the RAP1 signaling pathway enriched by DEGs described in Fig. 4a. e mRNA expressional correlation of TTC17 with members of the RAP1/CDC42 cascade in breast cancer, including RAP1A, RAP1B, CDC42, MAP2K6, RHOA, ITGB1, ITGB2, ACTB, and ACTG1. f, g Western blot analysis of RAP1/CDC42 signaling activation in MDA-MB-231 cells with or without TTC17 knockdown by increased RAP1A, CDC42 (f), and RAP1A-GTP (g). h, i Representative images and quantitative analysis of wound healing assays (h) and Transwell invasion assays (i) using TTC17-silenced MDA-MB-231 cells treated with the CDC42 inhibitor ML141 and their counterparts. Scale bars, h 500 μm; i 200 μm. j Representative immunofluorescence images of actin cytoskeleton and Golgi morphology in MDA-MB-231 cells with TTC17 silencing or scramble control. Scale bar, 10 μm. *P < 0.05, **P < 0.01, ***P < 0.001. DEGs, differentially expressed genes; FDR, false discovery rate; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; BRCA, breast invasive carcinoma