Fig 2: miR-494-3p regulates PTEN/Akt/NF-?B signaling. (A) miR-494-3p target prediction using four online tools. (B) Alignment of miR-494-3p and the 3'-UTR of PTEN. (C) Luciferase activity in 293T cells co-transfected with a reporter vector containing the 3'-UTR of PTEN and miR-494-3p mimics. RAW264.7 cells were transfected with miR-494-3p mimics, miR-494-3p inhibitor, NC or NC-i. After 24 h, cells were treated with 1 µg/ml LPS for 24 h. *P<0.05 vs. GLO-PTEN-WT. (D) PTEN expression was detected by confocal microscopy. (E) Western blot analysis was used to determine the protein expression levels of PTEN, Akt1, p-Akt1 and p65. (F) RAW264.7 cells were transfected with PTEN siRNA or control siRNA. After 48 h, cells were treated with 1 µg/ml LPS for 24 h. Western blot analysis was used to determine the protein expression levels of IL-1ß, TNF-a, PTEN, Akt1, p-Akt1 and p65. (G and H) Quantification of the protein expression presented in E and F. Data are presented as the mean ± standard deviation of at least three independent experiments. *P<0.05 (miR-494-3p) vs. nc; *P<0.05 (inhibitor) vs. NC-i; *P<0.05 (si-PTEN) vs. NC. Akt1, AKT serine/threonine kinase 1; IL, interleukin; LPS, lipopolysaccharide; miR, microRNA; MT, mutant; NC, negative control; NC-i, inhibitor NC; p, phosphorylated; PTEN, phosphatase and tensin homolog; siRNA, small interfering RNA; TNF, tumor necrosis factor; UTR, untranslated region; WT, wild-type.

Fig 3: mRNA expression of CASP3, TNF, PTGS2, TP3, NOS3 and VEGFA. Notes: ?? p < 0.01, ? p < 0.05 vs. Sham group; **p < 0.01, *p < 0.05 vs. I/R group (n = 3).

Fig 4: Protein expression of CASP3, TNF, PTGS2, TP3, NOS3 and VEGFA. Notes: ?? p < 0.01 vs. Sham group; **p < 0.01, *p < 0.05 vs. I/R group. 1: Sham, 2: I/R, 3: NLXTD5 g/ml, 4:NLXTD2.5 g/ml, 5: NLXTD1 g/ml, 6: NMDP (n = 3).

Fig 5: (A, B) Western blot analyses of lysates of p-p65/p65, MCP-1, TNF-a, IL-1ß; from Nrf2-/-+STZ+Resveratrol, Nrf2-/-+STZ+Vehicle and WT+STZ+Resveratrol mice using the indicated antibodies at 12 weeks after STZ injection(n = 3 mice per group; data are presented as means ± SEM).