Fig. 5
PF-271 treatments mimicked the osteoblast-induced dormancy in C4-2B cells. (A). Top list of AI-predicted potential dormancy-mimicking drugs based on the dormant gene signature of C4-2B cells in the mixed co-culture. C4-2B cells were treated with 1 µM/ 3 µM of PF-271 (PF) or Vehicle (Veh, DMSO) for 72 h. Cell proliferation monitoring by Incucyte (B), cell cycle phase distribution by PIP-FUCCI (C), Time-lapse NR2F1 protein expressions by immunoblotting (D), and mRNA expressions of NR2F1/MKI67/CCND1 (E) or MT-ND1/MT-CO1/MT-ATP6 (G) by qRT-PCR, were performed in the same manner as described in Fig. 2. (F). Analyses of apoptotic cells in C4-2B cells treated with 1 µM/ 3 µM of PF-271 (PF) or Vehicle (Veh, DMSO) for 72 h. H2O2 treatment for positive control. Scale bars, 50 μm (C) and 100 μm (F). All the in vitro experiments were independently repeated at least three times. The data in curves and bar plots are presented as the mean ± SD of each set of triplicate samples. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, and n.s. non-significant