Fig. 6: Spry2 mediates the effects of miR-21 on pancreatic cancer cells.
From: miR-21 promotes EGF-induced pancreatic cancer cell proliferation by targeting Spry2
a After being serum-starved for 24 h, MIA PaCa-2 cells transfected with EV or Spry2 plasmid were incubated in the absence or presence of 50 ng/ml EGF for the indicated times. Western blotting analysis of the indicated proteins was then carried out. Western blotting expression densitometry data was quantified by imageJ and then was normalized to the corresponding densitometry of GAPDH. b After being serum-starved for 24 h, PANC-1 cells were transfected with control siRNA (si-NC) or siRNA targeting Spry2 (si-Spry2) and incubated in the absence or presence of 50 ng/ml EGF for the indicated times. Western blotting analysis of the indicated proteins was then carried out. Western blotting expression densitometry data was quantified by imageJ and then was normalized to the corresponding densitometry of GAPDH. c miR-21 overexpressing PANC-1 cells transfected with EV or Spry2 overexpression plasmids were incubated in the presence of 50 ng/ml EGF. Then, CCK-8 proliferation assay was used to analyze the growth of PANC-1 cells. d miR-21 knockdown MIA PaCa-2 cells transfected with si-NC or si-Spry2 were incubated in the presence of 50 ng/ml EGF. Then, CCK-8 proliferation assay analyzed the growth of MIA PaCa-2 cells. e, f EdU assay further confirmed the proliferative activity of PANC-1 cells and MIA PaCa-2 cells. g Spry2 overexpression weakened the phosphorylation level of indicated proteins in miR-21 overexpressing PANC-1 cells after 50 ng/ml EGF treatment for 20 min. Western blotting expression densitometry data was quantified by imageJ and then was normalized to the corresponding densitometry of GAPDH. (H)Spry2 knockdown amplified the phosphorylation level of indicated proteins in miR-21 knockdown MIA PaCa-2 cells after 50 ng/ml EGF treatment for 20 min. Western blotting expression densitometry data was quantified by imageJ and then was normalized to the corresponding densitometry of GAPDH. Three independent experiments were performed for each group. All data are represented as the mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001
