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. 2019 Jun 16;16(10):1355–1363. doi: 10.1080/15476286.2019.1629769

Figure 4.

Figure 4.

DUSP3 downregulation reduces cell invasion and colony forming ability in A375 cells and alters MAPK/PI3K signalling. A) Western blot showing downregulation of DUSP3 protein in A375/211 cells and validating knockdown of DUSP3 upon transient DUSP3 siRNA (siDUSP3; 48 h post-transfection) transfection in A375 background compared to control siRNA (siNEG). B) DUSP3 knockdown does not affect A375 cell proliferation in vitro (right, compared to control siRNA, siNEG, based on four independent experiments). C) DUSP3 knockdown in A375 leads reduces cell invasion (right two panels; compared to control siRNA, siNEG). Reduced cell invasion is also seen in A375/211 compared to A375 cells (left two panels). Bar graph quantifying three independent trials. D) DUSP3 knockdown in A375 cells reduces colony formation (right two panels; compared to control siRNA, siNEG). Reduced colony formation is also seen in A375/211 compared to A375 cells (left two panels). Bar graph quantifying three independent trials. E) Human MAPK array depicting levels of various phosphorylated kinases with 8-minute exposure time. Boxed dots represent duplicate spots for identifying the indicated phosphorylated protein kinases. Bar graph representing normalized levels of selected (boxed) phosphorylated protein kinases for comparison between A375 vs. A375/211 and siNEG (control siRNA) and siDUSP3 (DUSP3 siRNA) conditions. F) Model of DUSP3 and miR-211-dependent dysregulation of MAPK and PI3K signalling. Significance level: * p < 0.05; ** p < 0.005; *** p < 0.001.