Fig. 3: The cleavage of GSDMD and proIL-1β in ASCs do not rely on the caspases of ASCs.

a ASCs were pretreated with VX765 or zVAD of indicated concentrations for 0.5 h before being primed with 0.5 μg/ml LPS for 4 h. Then ASCs were stimulated by SLNB mixed with VX765 or zVAD of indicated concentrations for 2 h. After the above treatments, replace the supernatant with fresh media for another 2 h-incubation or collect cells for immunoblot analyses (b, e) of indicated proteins. Data are representative of at least three independent experiments. c, f LDH release level of ASCs in the medium of a. n = 3. d, g ELISA detection of mature IL-1β in the media of a. n = 3. h–j LPS-primed WT and caspase-1/11^−/− ASCs were treated with SCB and SLNB for 2 h. Then replace the supernatant with fresh media for another 2-h-incubation or collect cells for immunoblot analyses (out of 3 performed) h of indicated proteins. LDH (i) and mature IL-1β (j) were detected in the medium. n = 3. Statistical significance was determined using one-way analysis of variance. Data are shown as mean ± SD. ns: p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. zVAD z-VAD-FMK, LPS lipopolysaccharide, ASCs adipose tissue-derived mesenchymal stem cells, BMDMs Bone-marrow-derived macrophages, SN supernatant, GSDMD gasdermin D, GSDMD-NT N-terminal gasdermin D, SLNB supernatant collected from LPS/NIG-treated BMDMs, NT no treatment of inhibitor, DMSO dimethyl sulphoxide, LDH lactate dehydrogenase, ELISA enzyme-linked immunosorbent assay.