Skip to main content
. 2019 Jun 12;10:360. doi: 10.3389/fendo.2019.00360

Figure 6.

Figure 6

CHRDL1 overexpression enhanced BMP-4-induced osteoblast differentiation in vitro. (A,B) CHRDL1 mRNA expression and protein levels were detected at 72 h after transfection of pLVX-CHRDL1. ***P <0.01 compared with pLVX-vector. (C) The mRNA levels of COL1A1, ALP, OCN, and OPN were detected at 72 h after pLVX-CHRDL1 transfection and 48 h after osteogenic induction. *P <0.05; ***P <0.01 vs. pLVX-vector transfected sample; and #P <0.01 vs. rhBMP-4 administrated sample. (D,E) ALP and Alizarin Red staining after transfection with pLVX-CHRDL1 and rhBMP-4 addition separately or in combination when cultured in osteogenic induction medium for 14 days. Data are presented as mean ±SD (n = 3). *P <0.05; ***P <0.01 vs. pLVX-vector transfected sample; and #P <0.001 vs. rhBMP-4 administrated sample. (F) Western blot analysis of BMPR II, p-Smad1/5/9, total Smad1/5/9, Runx2, CHRDL1, and GAPDH at 48 h after transfection with pLVX-CHRDL1 and rhBMP-4 addition separately or in combination. GAPDH was used as loading control. (G) mRNA levels of ALP, COL1A1, OCN, and OPN were detected 48 h after treating with LDN-193189 or its vehicle (DMSO). Data were presented as mean ± SD (n = 3); (*P <0.05 and #P <0.01). All experiments were repeated independently in triplicate.