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Fig. 2 | BMC Nephrology

Fig. 2

From: Resveratrol attenuates constitutive STAT3 and STAT5 activation through induction of PTPε and SHP-2 tyrosine phosphatases and potentiates sorafenib-induced apoptosis in renal cell carcinoma

Fig. 2

Pervanadate reverses the phospho-STAT3 inhibitory effect of RES. a and b Caki-1 and 786-O cells (1 × 106 cells/well) were incubated at 37 °C with various indicated concentrations of RES for 6 h. Whole-cell extracts were prepared, then equal amounts of lysates were analyzed by Western blot analysis using antibodies against p-JAK1(Tyr1022/1023), JAK1, p-JAK2(Tyr1007/1008), JAK2, p-Src(Tyr416), and Src. The results shown here are representative of three independent experiments. c Pervanadate reverses the p-STAT3 inhibitory effect of RES. Caki-1 and 786-O cells (1 × 106 cells/well) were co-incubated with the indicated concentrations of pervanadate and 50 μM RES for 6 h, after which whole-cell extracts were prepared and 15 μg portions of those extracts were resolved on 8 % SDS-PAGE gel, electrotransferred onto nitrocellulose membranes, and probed for p-STAT3(Tyr705) and STAT3. d Caki-1 and 786-O cells (1 × 106 cells/well) were treated with various indicated concentrations of RES for 6 h, after which whole-cell extracts were prepared and 10 μg portions of those extracts were resolved on 8 % SDS-PAGE, electrotransferred onto nitrocellulose membranes, probed for PTPε and SHP-2 antibody. The same blots were stripped and reprobed with β-actin antibody to verify equal protein loading. e Caki-1 and 786-O cells (1 × 106 cells/well) were treated with various indicated concentrations of RES for 6 h, and total RNA was extracted and examined for expression of PTPε C, PTPε M, and SHP-2 by RT-PCR. GAPDH was used as an internal control to show equal RNA loading. f Effect of PTPε knockdown on RES induced expression of PTPε. Caki-1 cells were transfected with either PTPε siRNA or scrambled siRNA (50 nM). After 48 h, cells were treated with 50 μM RES for 6 h and whole-cell extracts were subjected to Western blot analysis (left panels). Effect of SHP-2 knockdown on RES induced expression of SHP-2. 786-O cells were transfected with either SHP-2 siRNA or scrambled siRNA (50 nM). After 48 h, cells were treated with 50 μM RES for 6 h and whole-cell extracts were subjected to Western blot analysis (right panels)

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