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

Figure 2

Effect of RIPK1 on cell death of bone marrow MSCs. (a) Representative graphs were analyzed using the flow cytometer. After 72 h transfection with siRNA in bone marrow MSCs, Annexin V/PI staining was used to determine the type of cell death. Annexin V-positive and PI-negative represented apoptotic cells, while PI-positive represented necrotic cells. (b) Histogram analysis illustrated the increase of apoptotic and necrotic cells in RIPK1-deficient bone marrow MSCs. Values are expressed as the from three independent experiments (, vs. siControl; ). (c) Typical TEM images of morphological ultrastructural changes in bone marrow MSCs. After transfection with siRIPK1 for 72 h, bone marrow MSCs displayed both apoptotic ultrastructural changes including condensed nuclei and apoptotic body and necrotic ultrastructural changes including organelle swelling, plasma membrane disruption, and cellular lysis ( μm or 2 μm). (d) Representative western blots for the expression of Cleaved caspase-3, p-RIPK3, p-MLKL, and corresponding total protein. (e) Histogram analysis showing the relative protein levels of Cleaved caspase-3, p-RIPK3, p-MLKL, and corresponding total protein. Data are presented as the from three independent experiments (, vs. siControl; ).

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