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. 2021 Feb 8;6(3):e137245. doi: 10.1172/jci.insight.137245

Figure 5. Conversion of OS cells to adipocyte-like cells through TNIK inhibition.

Figure 5

(A–C) U2OS cells were treated with DMSO, 1 μM NCB-0846, or 1 μM NCB-0970 for 3 days, and intracellular lipid droplets were visualized by (A) Oil red O staining, (B) electron microscopy (EM), and (C) fluorescent molecular probe BODIPY labeling. Nc, nucleus. Arrowheads indicate lipid droplets. Scale bar: 20 μm (A and C); 3 μm (B). (D) U2OS cells were treated with DMSO, 1 μM NCB-0846, or 1 μM NCB-0970 for 3 days, and expression of the PPARG (n = 3, respectively) and FABP4 (n = 4 for DMSO and n = 6 for NCB-0846 and NCB-0970) genes (normalized to ACTB) was quantified by RT-PCR. Data represent the mean ± SD. One-way ANOVA, **P < 0.01, ****P < 0.0001. (E) Expression of the PPARG (n = 3) and FABP4 (n = 6) genes (normalized to ACTB) in U2OS cells stably expressing control shRNA or shTNIK was quantified by RT-PCR. Data represent the mean ± SD. One-way ANOVA, **P < 0.01. (F) Representative images of the macroscopic appearance of NOS-10 xenografts resected from mice administered vehicle or NCB-0846 HCl (90 mg/kg) for 10 days. Scale bar: 5 mm. (G) Oil red O staining for NOS-10 xenografts resected from mice administered vehicle or NCB-0846 HCl (90 mg/kg) for 10 days. Scale bar: 50 μm. (H) Expression of the PPARG and FABP4 genes (normalized to ACTB) in NOS-10 xenografts resected from mice administered vehicle (n = 9) or NCB-0846 HCl (n = 5) for 10 days. Student’s t test, *P < 0.05. Data represent the mean ± SEM.