Fig 1: Effects of JTE013 on cell growth and S1PRs expressions. Murine BMSCs were cultured in osteogenic media for 1 to 3 days with the presence of either DMSO or JTE013 (0.5 to 8 µM). Cell growth at (A) 24 h, (B) 48 h, and (C) 72 h was quantified by a CellTiter 96® AQueous One Solution Cell Proliferation Assay. (D) Relative S1PRs’ mRNA levels in murine BMSCs cultured in DMEM media with 10% FBS and 100 U/mL penicillin and streptomycin. (E–I) Murine BMSCs were treated with either DMSO or JTE013 (0.5 to 8 µM) and cultured in osteogenic media for 7 days. (E) S1PR1, (F) S1PR2, (G) S1PR3, (H) S1PR4, and (I) S1PR5 mRNA levels were quantified by RT-qPCR and normalized by GAPDH expression. (n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001). (J–O) Murine BMSCs were treated with either DMSO or JTE013 (0.5 to 8 µM) and cultured in osteogenic media for 9 days. (J) S1PR2, S1PR1, S1PR3, S1PR4, S1PR5, and GAPDH protein levels were measured by Western blot. Protein densitometry of (K) S1PR2, (L) S1PR1, (M) S1PR3, (N) S1PR4, and (O) S1PR5 was normalized by GAPDH protein expression in BMSCs (n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001).