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. 2021 Jun 10;11:12287. doi: 10.1038/s41598-021-91292-2

Figure 5.

Figure 5

HGK regulates ruffle formation and cancer cell motility in response to TNF-α. (A) Representative Western-blot analysis of the phosphorylated levels of JNKs and p38 proteins normalized with β-actin on the indicated PC3 cells. Serum-starved cells (for 16 h) were stimulated with TNF-α for 15 min or maintained untreated. Densitometric quantification are shown (B) Migration analysis of the indicated PC3 cell using TNF-α as chemoattractant. Left panels, representative images of migrating cells (bars: 100 µm); right panels, histograms showing the mean value ± S.E.M. of the number of migrating cells (n = 3). (C) Immuno-fluorescence microscopy images of phalloidin staining (red) in NTC and HGK depleted PC3 cells. Serum starved cells were stimulated with TNF-α for 15 min. Cell nuclei were stained with DAPI (blue). Scale bars: 150 and 75 μm. Right panel, histograms showing the mean value ± S.E.M. of cells with ruffles referred to non-silenced cells (n = 3). (D) Representative western-blot analysis of phosphorylated levels of JNKs and FAK-Y397 normalized with β-actin. Serum-starved PC3 cells (for 16 h) were stimulated with TNF-α for 15 min or maintained untreated. Full-length blots are presented in Supplementary Figure 3.