Fig. 4

Assessment of proliferation and apoptosis in human umbilical artery smooth muscle cells (HUASMCs) following manipulation of A2M expression and human decidua basalis. a, b, b1–b2 Determination of HUASMC viability in the control, A2M-overexpression and A2M-downregulated groups after 12-, 24-, 48-, 72-, and 96-h incubation by CCK-8 assay (a). Western blotting data showing the expression of A2M and α-SMA in HUASMCs (b), and b1–b2 show the quantitative analysis of A2M (b1) and α-SMA (b2) expression among the control, A2M-overexpression, and A2M-downregulated groups. c–f Flow cytometry data showing the analysis of the DNA contents in HUASMCs transfected with negative control (control) (c), A2M-silencing vector (A2Msi) (d), or A2M-overexpressing vector (A2M) (e), and f shows the quantitative analysis of the proportion of cells in each phase of the cell cycle in the three groups. g–j Apoptosis of HUASMCs transfected with negative control (control) (g), A2M-silencing vectors (h), or A2M-overexpression vector (i) was determined by flow cytometry using the annexin V-FITC/PI apoptosis assay, and j shows the quantitative analysis of the cell apoptosis rates in the three groups. k, k1 Representative double immunofluorescence staining of α-SMA and PCNA on the cross-sections of the spiral arteries (counterstained with DAPI) of the third trimester from the normal and PE groups, and k1 shows the quantitative analysis of PCNA expression in both groups. l, m Western blotting data showing the expression of PCNA (l) and FAS (m) and the quantitative analysis of the human decidua basalis of the third trimester from the normal and PE groups. Scale bars = 50 μm in k. *P < 0.05, **P < 0.01, ***P < 0.001