Fig 2: Knockdown of HOXC10 suppresses the WNT-EMT process in OSCC cell lines. (A). FaDu cells and (B) SCC4 cells were treated with shHOXC10, and Wnt10B, N-cadherin, E-cadherin, and Vimentin levels were determined. GAPDH served as an internal standard for protein loading. (C). FaDu cells and (D) SCC4 cells were treated with negative control (NC) and shHOXC10; representative immunofluorescence is shown, and fluorescence of N-Cadherin, Snail and E-cadherin was quantified; scale bar: 20 µm. The data are presented as the means ± SEM. **P<0.01 versus the control group.

Fig 3: PADI4 was involved in the process of EMT. (A) Knockdown of PADI4 increased the expression of epithelial makers (E-cadherin) and reduced the expression of mesenchymal markers (N-cadherin and vimentin). (B) The statistical chart represents the relative protein expression levels. (C) Knockdown of PADI4 reduced the EMT-associated Snail1/Smad3/4 transcriptional factors compared with the control group. (D) The statistical analysis represents the relative protein expression levels. Differences among groups were analyzed by one-way analysis of variance, followed by Bonferroni's multiple comparison test. *P<0.05 or **P<0.01 vs. control, n=3. N, neural; E, epithelial; EMT, epithelial-mesenchymal transition; PADI4, protein-arginine deiminase type-4; Smad, mothers against decapentaplegic homolog; Ctrl, control; NC, negative control.

Fig 4: Prevalence of EMT-like phenotype in the RPE cells of human AMD tissues. RPE cell lysates from human AMD samples showed increased Snail1 and vimentin expression concomitant with decreased expression of E-cadherin compared to age-matched controls. Representative Western blot images are shown. The graphs show mean ± SD and *P < 0.05 (Mann-Whitney U test), n = 4.

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.