Fig. 5

hUMSCs reduced LPS + ATP-induced BV2 microglia pyroptosis by secreting TSG-6. BV2 cells were co-cultured with hUMSCs, sh-TSG-6 hUMSCs, or sh-NC hUMSCs activated by TNF-α and different concentration of rmTSG-6 (50 ng/mL, 100 ng/mL, 200 ng/mL) and treated with or without LPS (1 µg/mL) + ATP (5 mM) for 12 h. a Representative photomicrographs and quantification of double staining with PI (red) and DAPI (blue) were captured by confocal microscopy. b Caspase-1 activity and the release of LDH in the supernatant was assessed using Caspase-1 activity and LDH activity detection kits. c The protein levels of IL-1β, pro IL-18, and TNF-α were evaluated by ELISA. d Representative protein bands and corresponding grayscale values of NLRP3, pro-Caspase-1, Caspase-1 p20, GSDMD and cleaved-GSDMD were performed by western blotting. Scale bar = 100 μm. All data are represented as means ± SD of at least 3 independent experiments and compared by one-way ANOVA, followed by the Tukey’s post hoc test. *p < 0.05, **p < 0.01, versus LPS + ATP group; ^#p < 0.05, sh-TSG-6 hUMSC group versus sh-NC hUMSC group