Fig. 7

sTNFRII-Fc modification protects MSCs against apoptosis, autophagy and immunogenicity induced by TNF-α and restores the immunosuppressive capacity of MSCs. a, b Cells were incubated with MSC-CM, EGFP-MSC-CM or sTNFRII-MSC-CM in the presence of TNF-α + CHX as indicated, and stained with annexin V-FITC and PI-PE, and then analyzed by flow cytometry (b) and representative fluorescent images were collected by a imaging flow cytometer (a). c The percentage of apoptotic cells were analyzed (n = 5). d Cells treated with/without TNF-α + CHX, and the expression levels of Bcl-2, Bax, caspase-3, cleaved caspase-3, caspase-8, cleaved caspase-8, LC3B-II and TRIB3 were assessed by western blot. e Densitometry quantification of western blot from (d). The intensity of each was normalized to β-actin intensity (n = 3). f sTNFRII-Fc modification restores the immunosuppressive capacity of MSCs. Cells were treated as indicated, and the cell viability was measured by CCK-8 assay (n = 3). g The expression of HLA-DR on sTNFRII-MSC stimulated with TNF-α and IFN-γ was determined by flow cytometry. h Quantification of the fluorescence intensity from (g) (n = 3). Data are presented as the mean ± SD values. *p < 0.05, **p < 0.01