Fig 2: miR-3656 expression modulates the gemcitabine sensitivity of PC cells in vivo. (a) Tumor growth curves of PANC-1 cells transfected with a miR-3656 construct or empty vector and treated with gemcitabine or saline. (b and c) Representative images showing the tumors formed in the four groups, including control overexpression + saline (OE-Con + Saline), miR-3656 overexpression + saline (OE-miR-3656 + Saline), control overexpression + gemcitabine (OE-Con + Gemcitabine), miR-3656 overexpression + gemcitabine (OE-miR-3656 + Gemcitabine), at the 5th week after subcutaneous transplantation or when mice were killed. (d) Comparison of the mean tumor weights of the four groups. (e) Representative IHC images for RHOF, TWIST1, E-cadherin and Vimentin or TUNEL staining among the four tumor xenografts groups. (f-j) Statistical comparisons of RHOF, TWIST1, E-cadherin, Vimentin expression and apoptotic cell numbers among the four tumor xenografts groups. All n=6; bar, S.E.M., n.s, not significant, *P<0.05; **P<0.01; ***P<0.001; Student’s t-test

Fig 3: Immunohistochemistry. P-STAT3, ZEB1, and Twist1 in squamous cell carcinoma tissues and adjacent tissues were detected by immunohistochemistry. P value was written in the figure.

Fig 4: Effect of diabetes and Akt2 cKO on the expression of EMT markers in RPE.8 months of diabetes (10 months of age) significantly increased the mRNA levels of EMT markers Snai1 (A), Snai2 (B), and Twist1 (C) but not for Twist2 (D) in the RPE cells from Akt2fl/fl diabetic mice; Akt2 cKO inhibited this diabetes-induced increase. n = 6 mice for each group. Data are shown as mean ± SD. ***p < 0.001 and ****p < 0.0001. Statistical test used in this study is One-way ANOVA followed by Tukey’s post-hoc test. N non-diabetic, D diabetic, cKO conditional knock-out.

Fig 5: Akt2 cKO attenuates diabetes-induced increased protein level of EMT markers and decreased tight junction proteins in RPE.A Representative RPE immunoblots and quantification of protein levels of EMT markers B Snail, C Twist1, D Vimentin, E adherens junction protein E-cadherin and F, G tight junction proteins ZO-1 and Occludin from 10 month old (8 months of diabetes) mice in Akt2fl/fl and Akt2 cKO RPE tissue. Diabetes significantly increased the levels of Snail, Twist1, and Vimentin, and decreased the levels of E-cadherin, ZO-1, and Occludin in the RPE cells from diabetic Akt2fl/fl mice compared to non-diabetic controls. However, Akt2 cKO significantly inhibited these diabetes-induced effects. n = 6 mice per group. Data are shown as Mean ± SD. *p < 0.05 and **p < 0.01. Statistical test used in this study is One-way ANOVA followed by Tukey’s post-hoc test. N non-diabetic, D diabetic, cKO conditional knock-out.