RIPK1 Coordinates Bone Marrow Mesenchymal Stem Cell Survival by Maintaining Mitochondrial Homeostasis via p53

Figure 4

RIPK1 regulated proliferation and differentiation of bone marrow MSCs via p53. (a) Representative western blots for the expression of p53. p53 expression was upregulated in bone marrow MSCs following RIPK1 inactivation. Histogram analysis showing the relative protein levels of p53. Data are presented as the from three independent experiments (, vs. 0 h group; ). (b) The bone marrow MSCs were transfected with sip53 for 24 h; then, the knockdown efficiency of p53 was assessed by western blotting. Representative western blots of the expression of p53. Histogram analysis showing the relative protein levels of p53. Data are presented as the from three independent experiments (, vs. siControl; ). sip53(#1) showed the highest efficiency among three sip53. (c) The inhibition effect of siPIRK1 on cell proliferation was significantly reversed by sip53, by CCK-8 assay. Values are expressed as the from three independent experiments (, vs. siRIPK1; ). (d) Alizarin Red staining showed the inhibition of siRIPK1 on osteogenic differentiation was reversed by sip53, and (e) Runx2 expression level in siRIPK1 was increased after transfected with sip53. Values are expressed as the from three independent experiments (, vs. siRIPK1; ) ( μm). (f) Oil Red O staining showed the inhibition of siRIPK1 on adipogenic differentiation was reversed by sip53, and (g) PPARγ expression level in siRIPK1 was increased after transfected with sip53. Values are expressed as the from three independent experiments (, vs. siRIPK1; ) ( μm). (h) Alcian Blue staining showed the inhibition of siRIPK1 on chondrogenic differentiation was reversed by sip53, and (i) Sox9 expression level in siRIPK1 was increased after transfected with sip53. Values are expressed as the from three independent experiments (, vs. siRIPK1; ) ( μm).

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