Research Paper Volume 15, Issue 14 pp 6834—6847

Figure 3. ATF4 knockdown delays the osteogenic differentiation of VICs. (A) IF staining of α-SMA and vimentin proteins in the VICs isolated from non-calcified valve tissues. (B) RT-qPCR detection of BMP2, OPN, and osteocalcin expression in the VICs induced by OIM and normal culture medium. (C) ALP activity in the OIM-induced VICs. (D) RT-qPCR detection of ATF4 expression in the VICs induced by OIM. (E) Western blot analysis of ATF4 protein in the OIM-induced VICs. (F) RT-qPCR detection of ATF4 expression in the OIM-induced VICs treated with sh-ATF4. (G) RT-qPCR detection of BMP2, OPN, and osteocalcin expression in the OIM-induced VICs treated with sh-ATF4. (H) ALP activity in the OIM-induced VICs treated with sh-ATF4. ^*p < 0.05. Cell experiments were repeated three times independently. Measurement data were expressed as mean ± standard deviation. Data obeying normal distribution and homogeneity of variance between two groups were analyzed using unpaired t-test.