FIGURE 6.

SIRT1 promotes trophoblast EMT. (a) Venn diagram of SIRT1‐binding proteins in HTR8/SVneo cells and human first‐trimester villi profiled by IP‐MS. (b) Reciprocal Co‐IP of SIRT1 and vimentin in human first‐trimester villi (left panel) and HTR8/SVneo cells (right panel). (c) Acetylation of vimentin in WT and shSIRT1 HTR8/SVneo cells. (d) Western blot analysis of E‐cadherin, N‐cadherin, vimentin, and SNAIL in WT, shNC, and shSIRT1 HTR8/SVneo cells. (e) Acetylation of vimentin in Elf5 ^cre/+, Sirt1 ^fl/fl and Sirt1 ^fl/fl mouse placentas collected on E13.5. (f) Western blot analysis of E‐cadherin, N‐cadherin, vimentin, and SNAIL levels in Elf5 ^cre/+, Sirt1 ^fl/fl and Sirt1 ^fl/fl mouse placentas collected on E13.5; n = 3. (g) Acetylation of vimentin in first‐trimester villi of AMA and control pregnancies. (h) Western blot analysis of E‐cadherin, N‐cadherin, vimentin, and SNAIL levels in first‐trimester villi from women with AMA and control women; n = 3. AU, arbitrary unit. Data were analyzed using Student's t‐test; *p < 0.05, **p < 0.01. All data are presented as the mean ± SEM. All experiments were performed in triplicate. (i) Transwell assays of Htr8/SVneo cells were performed after 48 h of treatment with siVim, SRT1720 or both. (j) Western blot analysis of E‐cadherin, N‐cadherin, vimentin, and SNAIL levels in Htr8/SVneo cells treated with siVim, SRT1720, or both for 48 h. Data were analyzed using one‐way ANOVA; **p < 0.01, ***p < 0.001. All data are expressed as the mean ± SEM