The change of inflammsomes in BA-MSCs after priming with 66PR in vitro.
A small-molecular compound, 66PR, was synthesized, characterized, and used to treat BA-MSCs. Then the cells were primed for 4 h with LPS and 66PR (20 μM) and stimulated with or without nigericin for 1.5 h before the following observations. (a) Synthesis of 66PR. (b) Scanning electron microscopic observation of BA-MSCs treated with 66PR. (c)Analysis of pore number counts of BA-MSCs after 66PR and mock treatment according to (b). (d) Quantity of GSDMDC1+ foci from BA-MSCs after 66PR and mock treatment based on confocal laser scanning microscopic analysis. (e) ELISA analyses of IL-1β in BA-MSCs supernatant after 66PR and mock treatment. (f) Western blot analysis of NLRP3, caspase-1, and caspase-11 in wild type BA-MSCs after 66PR and mock treatment. (g) Western blot analysis of caspase-1 and IL-1β in caspase-11-/-BA-MSCs after 66PR treatment; (h) Western blot analysis of caspase-11 in NLRP3-/-BA-MSCs after 66PR treatment. CL: cell lysates, SN: supernatants. At least 300 cells were counted for morphological analyses, mean ± SD, two-way ANOVA, *P < 0.05,**P < 0.01,***P < 0.001, ns: not significant, nd: no detected.