Fig. 5

Involvement of macropinocytosis in extracellular ATP-induced migration and EMT induction. a A549 cells were transfected with scrambled siRNA or sorting nexin 5 (SNX5) siRNA and incubated for 48 h. The expression of SNX5 was detected by Western blot. b 48 h after transfection, cells were incubated with 0, 0.5 or 1.0 mM ATP and intracellular ATP levels were measured by ATP assay. c, d 48 h after transfection, ATP-induced cell invasion was examined using Transwell assays (c), and the quantitative analysis of invasive cells (d). e Control and SNX5 knockdown cells were treated with 0, 0.5 or 1.0 mM ATP for 24 h before analysis. EMT-associated proteins: E-cadherin, vimentin, β-catenin and Snail protein expression levels were measured by western blot. f Densitometry analyses of individual blots were performed using ImageJ software. Experiments were performed in triplicate and data are presented as the mean ± standard deviation. *P < 0.05, **P < 0.01 and ***P < 0.001