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. 2020 Mar 4;10(9):3980–3993. doi: 10.7150/thno.41028

Figure 5.

Figure 5

TMEM16A ameliorated autophagy-meditated vascular remodeling by reducing VPS34 activity. (A-C) Western blot analysis of the phosphorylation of AKT, mTOR (A), p70S6K, and 4EBP1 (B) and the expression of VPS34 (C) in aortas from TMSMC Tg and TMcon mice following AngII infusion for 4 weeks. (D) Bar charts showing VPS34 kinase activity in aorta homogenates. **P < 0.01 vs. sham + TMcon; ##P < 0.01 vs. AngII + TMcon. Statistical significance was determined by one-way ANOVA. n = 6 mice/ group. (E) TMcon or TMSMC Tg mice were injected intraperitoneally with 3-MA (100 µg/kg) or rapamycin (1 mg/kg), respectively, at 2 weeks after AngII infusion, and then continuously dosed once per day for 2 weeks. Representative images of Ki67 staining and hematoxylin-eosin staining of aortic sections are shown. Scale bars, 25 µm (upper panel), 200 µm (middle panel) or 50 µm (lower panel). (F) Quantification of cross-sectional areas (CSAs). **P < 0.01 vs. sham + TMcon; ##P < 0.01 vs. AngII + TMcon; &&P < 0.01 vs. AngII + TMSMC Tg, one-way ANOVA. n = 6 mice/ group. (G) Cells were infected with Ad-Lacz or Ad-TM for 48 h and then transfected with VPS34 plasmid in the presence of AngII for 24 h. Cell proliferation was determined by performing BrdU assays. (H) Proliferation of MASMCs transfected with scr.RNA or TM siRNA following AngII and SAR405 (1 µmol/L) treatment for 24 h. **P < 0.01 vs. control; ##P < 0.01 vs. AngII; &&P < 0.01 vs. AngII + Ad-TM or AngII + TM siRNA, one-way ANOVA. n = 6.