The effects of KY19382 on chondrocyte proliferation and differentiation. The effects of KY19382 on chondrocyte proliferation and differentiation. (A) ATDC5 cells treated with 0.01 or 0.1 μM concentrations of KY19382 were incubated for 48 h followed by treatment with 50 μM BrdU for 12 h before harvesting. BrdU incorporation was visualized by immunofluorescent staining using a specific BrdU antibody (left). BrdU-positive cells were quantified (mean ± SEM, n = 3; ANOVA, P = 7.8 × 10−5; Tukey’s post-hoc test, *P < 0.05 and ***P < 0.0005) (right). Scale bar, 100 μm. (B) qRT-PCR analyses of mRNA levels of chondrogenic differentiation markers in ATDC5 cells incubated with 0.1 μM KY19382 for 3 d in three-dimensional alginate beads after transfection with control siRNA or Ctnnb1 siRNA (mean ± SEM, n = 3; t test, *P < 0.05, **P < 0.005, and ***P < 0.0005). (C) qRT-PCR analyses of mRNA levels of chondrogenic differentiation markers in C28/I2 cells incubated with 1 μM KY19382 for 3 d in three-dimensional alginate beads (mean ± SEM, n = 3; t test, *P < 0.05 and ***P < 0.0005). n.s., no significance. Sehee Choi et al. LSA 2019;2:e201800254 © 2019 Choi et al.