Fig. 2

AS-IV promotes mitochondrial function and proliferation of HSC in vitro. A BMNCs were isolated from KM mice and cultured in vitro. The cell viability of bone marrow nucleated cells was assessed using the CCK-8 assay on day 5 and 9 after administration of AS-IV intervention; B Bone marrow nucleated cells from eGFP-fluorescent mice were extracted and cultured in vitro, and their proliferation was counted by fluorescence photography and fluorescence intensity analysis after the intervention of AS-IV administration on day 5 and 9; C Cell colony formation assay to detect CFU-E, BFU-E, CFU-GM proliferation after AS-IV intervention on bone marrow nucleated cells; D–E Bone marrow nucleated cells treated with AS-IV-labelled fluorescent probe JC-1 (2.5, 5, 10 μM) to assess changes in mitochondrial membrane potential by inverted fluorescence microscopy. JC-1 polymer appears as red, whereas JC-1 monomer appears as green. Scale bar: 100 μm; F–G Mito-Tracker green fluorescent probe for detection of mitochondrial mass after drug administration. Nuclei were stained with Hoechst 33,258 (scale bar: 100 μm); H ATP Assay Kit detected the concentration of ATP produced by cells; I After 9 days of in vitro culture of bone marrow nucleated cells, the expression of c-kit⁺ HSC in each group was evaluated using flow cytometry; J Histograms showing the percentage of c-kit⁺ HSC in each group; K–L Mitochondrial membrane potential changes of c-kit⁺ HSC detected by flow cytometry. Data represent the mean ± standard deviation of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 compared with control