Fig. 7

miR-584-5p promoted cellular senescence and negatively regulated TGFβ signaling pathway via downregulation of WWP1. a. miR-584-5p up-regulation or WWP1-shRNA promoted cellular senescence by detecting SA-β-gal activity. The rescue experiments for miR-584-5p overexpression were performed by ectopic expression of WWP1 without its 3’-UTR in MGC803 cells. b. Similar rescue experiments for miR-584-5p silencing was performed by down-regulation of WWP1 in SGC7901 cells. c. The expression levels of senescence-related proteins were evaluated by Western blot. d. Expression of p53 signifcantly increased by miR-584-5p was ameliorated by the p53 inhibitor pifthrin-α. e. Treatment of pifthrin-α signifcantly receded miR-584-5p induced apoptosis of the MGC803 cells which was measured in a flow cytometer. f. Positive SA-β-Gal stained cells induced by miR-584-5p was confined by treatment of pifthrin-α. h. Levels of key proteins involved in TGFβ signaling by western blot. i. Levels of key proteins involved in TGFβ signaling in MGC803 cells treated with LY364947 were measured by western blot. j. The expression level of TGFβ was determined in MGC803 and SGC7901 by immunofluorescence assays. Scale bar is 25 μm. *p < 0.05, **p < 0.01, ***p < 0.001. The data expressed as the mean ± SD