Figure 4

hBcl2 overexpressing BMSCs with enhanced anti-apoptotic ability could resist myelin debris-induced early apoptosis in vitro. (A) Cellular immunofluorescence staining of cleaved caspase3 was used to assess apoptosis in the hBcl2, cb, NC, and BMSCs groups after incubating cells with macrophage conditioned medium (Mφ CM) for 24 h. White arrows represent apoptotic cells. (B) Cleaved caspase3 immunofluorescence staining was used to determine the apoptosis of cells in each group after 24 h cell culture in Dil-myelin CM (500 μg/mL). (C,D) Quantitative analysis of the proportion of cleaved caspase3 positive cells in the total number of cells, which were culture in macrophage CM (C) or myelin debris (D) (At least three independent replicates were performed for each experiment. Data are presented as mean ± SD, ****P < 0.0001 vs. hBcl2 group, cb vs. NC group, analyzed by one-way ANOVA, followed by Tukey’s post hoc test). (E) The morphological changes of apoptotic cells (black arrow) were observed under a light microscope after 24 h cell culture in myelin debris (500 μg/mL). Scale bars = 50 μm (A,B,E).