Fig. 3: MSCs-induced megakaryocytic differentiation through restoring MPL expressions of CML cells.

A K562 cells (left) and LAMA-84 cells (right) were co-cultured with MSCs, respectively. The expression levels of MPL were measured by RT-qPCR (n = 3). B K562 cells were co-cultured with MSCs for 48 h, the expression of upstream transcription factors of MPL in K562 cells was assessed by RT-qPCR (n = 3). C K562 cells were treated with MSCs for 48 h, the cell-surface expression of MPL was measured by western blotting. ATPase Na⁺/K⁺ beta 2 (ATPase) was used as a loading control for membrane fraction. D K562 cells were cultured alone or co-cultured with MSCs for the indicated time. MPL levels were measured by flow cytometric. E K562 cells were transfected with siRNA negative control (K562 si-NC) or MPL-siRNA (K562 si-MPL). The K562 si-NC or K562 si-MPL cells were co-cultured with MSCs. After 48 h, the cell surface protein CD41 and CD42b in K562 cells were analyzed by flow cytometric. F The phosphorylation status of correlated signaling pathways was determined by western blotting after treatment with MSCs for 48 h. GAPDH was used as a loading control. G The expression levels of THPO in MSCs were assessed by RT-qPCR (n = 3). H K562 cells were overexpressed MPL (OE-K562), the relative mRNA levels of differentiation genes were assessed by RT-qPCR after treatment with El for 48 h. I CD41 and CD42b levels of OE-K562 cells were measured by flow cytometric after treatment with El for 48 h. All statistical data in this figure represent the mean ± SEM (*P <0.05, **P <0.01, ***P <0.001, ns, not significant, by one-way ANOVA in A, D, and G by Student’s t test in B, C, E, H, and I).