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. 2018 May 26;32:31–42. doi: 10.1016/j.ebiom.2018.05.023

Fig. 1.

Fig. 1

The activation of NLRP3 inflammsome in BA-MSCs was induced by nigericin combined with LPS.

Wild type or NLRP3 gene knockout BA-MSCs were primed for 4 h with LPS (500 ng/ml) or not, and then stimulated with or without nigericin (5 μM) for 1.5 h, before the following observations. (a) Scanning electron microscopic observation of NLRP3−/-BA-MSCs and wild type BA-MSCs (n = 8). (b) Analysis of pore number counts of wild type BA-MSCs according to (a). (c) Comparison of pore number counts between wild type BA-MSCs and NLRP3−/-BA-MSCs according to (a). (d) Confocal laser scanning microscopic analysis of nuclei (DAPI, blue) and GSDMDC1 (red) foci of NLRP3-/-BA-MSCs and wild type BA-MSCs. (e) Quantity of wild type BA-MSCs containing GSDMDC1+ foci from (d). (f) Quantity of both BA-MSCs type containing GSDMDC1+ foci from (d). (g) Western blot analysis of NLRP3, caspase-1and IL-1β proteins in total lysates (CL) and supernatants (SN) of NLRP3-/- BA-MSCs and wild type BA-MSCs using the indicated antibodies. n = 3, pro-IL-1β: IL-1β precursor; IL-1β: mature IL-1β; pro-Casp1: caspase-1 precursor; Casp1p20: caspase-1p20 subset. (h-i) ELISA analysis of IL-1β/18 levels in the supernatants of wild type BA-MSCs, n = 5. (j-k) ELISA analysis of IL-1β/18 levels in the supernatants of NLRP3-/-BA-MSCs and wild type BA-MSCs, n = 5. At least 300 cells were counted for morphological analyses, mean ± SD, t-test, *P < 0.05,**P < 0.01,***P < 0.001, ****P < 0.0001, ns: not significant. Ctrl: control; LPS: BA-MSCs were stimulated by LPS; LPS + NIG: BA-MSCs were stimulated by LPS and nigericin.