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. 2020 May 28;7(13):1903451. doi: 10.1002/advs.201903451

Figure 5.

Figure 5

Small molecule OB‐4 could promote osteogenic differentiation and enhance ATF4 protein expression in vitro. a) ALP staining showing ALP activity in MC3T3‐E1 cells incubated with the small molecules OB‐1, OB‐2, OB‐3, and OB‐4 at 1 µm or DMSO for 48 h, respectively. b) Real‐time PCR analysis of osteocalcin mRNA level in MC3T3‐E1 cells after treatment with the small molecules OB‐1, OB‐2, OB‐3, and OB‐4 at 1 µm or vehicle for 48 h, respectively. c) Representative Western blot (left) and quantification (right) of ATF4 protein level in MC3T3‐E1 cells after treatment with the small molecules OB‐1, OB‐2, OB‐3, and OB‐4 at 1 µm or vehicle for 48 h, respectively. d) Real‐time PCR analysis of ATF4 mRNA level in MC3T3‐E1 cells after treatment with small molecules OB‐1, OB‐2, OB‐3, and OB‐4 at 1 µm or vehicle for 48 h, respectively. e) ALP staining showing ALP activity in MC3T3‐E1 cells incubated with the small molecule OB‐4 at a series of concentrations (40, 60, 80 nm) or DMSO for 48 h. f) Real‐time PCR analysis of osteocalcin mRNA level in MC3T3‐E1 cells after treatment with the small molecule OB‐4 at a series of concentrations (40, 60, 80 nm) or vehicle for 48 h. g) Representative Western blot (left) and quantification (right) of ATF4 protein level in MC3T3‐E1 cells after treatment with the small molecule OB‐4 at a series of concentrations (40, 60, 80 nm) or vehicle for 48 h. h) Real‐time PCR analysis of ATF4 mRNA level in MC3T3‐E1 cells after treatment with the small molecule OB‐4 at a series of concentrations (40, 60, 80 nm) or DMSO for 48 h, respectively. The data are presented as the mean ± standard deviation. *p < 0.05, **p < 0.01, ***p < 0.001 versus vehicle group.