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. Author manuscript; available in PMC: 2023 Jan 21.
Published in final edited form as: Circ Res. 2021 Dec 10;130(2):184–199. doi: 10.1161/CIRCRESAHA.121.318881

Fig. 2. Cholesterol accumulation and altered lysosomal function in human VSMCs Wnt5a−/−.

Fig. 2

Human VSMCs Wnt5a−/− and Wnt5a+/+ were analyzed untreated (D0) or upon treatment with a cholesterol accumulation protocol during 10 days (D10). A, Quantification of total cholesterol n=5 D0 Wnt5a+/+, n=7 D0 Wnt5a−/−, n=8 D10 Wnt5a+/+, n=10 D10 Wnt5a−/−). B, Quantification of cholesteryl esters (n=5 D0 Wnt5a+/+, n=7 D0 Wnt5a−/−, n=8 D10 Wnt5a+/+, n=10 D10 Wnt5a−/−). C, Relative transcript levels of SREBP-2 (n=4 D0 and D10) and HMG CoA reductase (n=9 D0 Wnt5a+/+, n=8 D10 Wnt5a−/−, n=4 D10 Wnt5a +/+ and n=4 D10 Wnt5a −/−) taking GAPDH 18S as internal control. D, Oil Red O staining upon treatment for cholesterol accumulation during 10 days in presence of human recombinant Wnt5a (+rWnt5a) or BSA (+BSA) in human Wnt5a−/− VSMCs (n=3). E, Representative electron micrographs show presence of large lipofuscin containing vesicles (Lipo), lipid droplets (LD), and large LELs in human Wnt5a−/− VSMCs vs controls. Scale bar 1μm (n=3). A, B, and C show individual value along with mean±SEM. Statistical significance was assessed by Shapiro-wilk to test normality, unpaired t test for A and B, and Mann-Whitney test for C. A, P=0.034; B, P=0.038; C, P=0.028 and P=0.028 for SREBP-2 and P=0.034 and P=0.028 for HMG CoA reductase indicate significance relative to Wnt5a+/+. ns indicates not significant.