Figure 4. ATF4 silencing represses AVC in mice. (A) ARS staining of aortic valve leaflet calcification in HC-induced mice treated with sh-ATF4. (B) Expression of ATF4 in the aortic valve tissue of HC-induced mice treated with sh-ATF4 detected by RT-qPCR. (C–E) Expression of BMP2, OPN, and osteocalcin in the aortic valve tissue of HC-induced mice treated with sh-ATF4 detected by RT-qPCR, IHC, and IF. (F) IHC of CD86 protein in the aortic valve tissue of HC-induced mice treated with sh-ATF4. *p < 0.05. n = 8. Measurement data were expressed as mean ± standard deviation. Data obeying normal distribution and homogeneity of variance among multiple groups were analyzed by one-way ANOVA with Tukey's post hoc tests.