Downregulation of TRPC4 and TRPC5 Inhibits Smooth Muscle Cell Proliferation without Affecting Endothelial Cell Proliferation

Figure 2

(a) siTRPC4 (100 nM) and siTRPC5 (100 nM) were transfected into VECs and VSMCs, and cells were examined 48 h after transfection by fluorescence microscopy. a and b are unlabeled Cy3 negative controls for transfection of VECs for 48 h; c and d are TRPC4 siRNA (100 nM) for transfection of VECs for 48 h, e and f are TRPC5 siRNA (100 nM) for transfection of VECs for 48 h; g and h are TRPC4 siRNA (100 nM) and TRPC5 siRNA (100 nM) which were cotransfected into VECs for 48 h. Among them, a, c, e, and g are observations of cell morphology after transfection under brightfield, b, d, f, and h are observations of transfection efficiency under the red fluorescence channel, i and j are unlabeled Cy3 negative controls for transfection of VSMCs for 48 h, k and l are TRPC4 siRNA (100 nM) for transfection of VSMCs for 48 h, m and n are TRPC5 siRNA (100 nM) for transfection of VSMCs for 48 h, and o and p are TRPC4 siRNA (100 nM) and TRPC5 siRNA (100 nM) which were cotransfected into VSMCs for 48 h. Among them, i, k, m, and o are observations of cell morphology after transfection under brightfield, and j, l, n, and p are observations of transfection efficiency under the red fluorescence channel. Scale bars = 50 μm. (b) After expressions of TRPC4 and TRPC5 were suppressed using an siRNA strategy, TRPC4 and TRPC5 protein expression levels were measured by western blotting. BLK: blank group. N = 12. vs. VSMC-CTRL, VSMC siTRPC4, and VSMC siTRPC5. .

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