Figure S3. DOCK1 promotes cell motility via the DUSP4-ERK signaling pathway.

(A) Volcano plot analysis showing the differentially expressed genes; genes with log₂ foldchange > 1 and P-value < 0.05 are indicated in red; genes with the log₂ foldchange < −1 and P-value < 0.05 are indicated in blue; genes with the |log₂ foldchange| < 1 or P-value > 0.05 are indicated in grey. (B) ERK protein phosphorylation levels in JAR cells with DOCK1 repression or overexpression. (C) ERK phosphorylation levels in HTR-8 cells treated with 10 ng/ml EGF for 0, 30, and 60 min. (D) Knockdown efficacy of DUSP4 siRNA and DOCK1 expression after DUSP4 knockdown detected in HTR-8 cells by Western blotting. (E) 293T cells were transfected with ectopic Flag-DOCK1, Myc-DUSP4, and HA-UB for 48 h, and then treated with 20 μM MG132 for 6 h. The cell lysates were immunoprecipitated with Myc antibody and ubiquitination assays showed the DUSP4 protein ubiquitination levels. (F) mRNA expression of DUSP4 after HUWE1 knockdown in HTR-8 cells. (G) Knockdown efficiency of HUWE1 siRNA and DUSP4 protein abundance after HUWE1 knockdown detected in HTR-8 cells by Western blotting.