Extended Data Fig. 8: Regulation of LPS-induced cytokine production by mtROS in BMDMs. | Nature Metabolism

Extended Data Fig. 8: Regulation of LPS-induced cytokine production by mtROS in BMDMs.

From: Pro-inflammatory macrophages produce mitochondria-derived superoxide by reverse electron transport at complex I that regulates IL-1β release during NLRP3 inflammasome activation

Extended Data Fig. 8

a, Mitochondrial superoxide production measured with MitoNeoD, ∆ψm measured with TMRM, and succinate and itaconate measured by mass spectrometry in nonstimulated (NS) BMDMs or BMDMs stimulated with LPS for 3–24 h (n = 6-9). Data are scaled to the relative minima and maxima of each measurement (MitoNeoD (n = 3 (NS, 6 h, 9 h), n = 6 (3 h, 18 h, 24 h)); ∆ψm (n = 6 (3 h, 6 h, 18 h, 24 h), n = 9 (NS, 9 h)); succinate (n = 3); itaconate (n = 3)). b, IL-10, TNFα and IL-6 release measured by ELISA from NS BMDMs or BMDMs stimulated with LPS for 3–24 h (n = 3). Data are mean +/- S.E.M. of biological replicates. b,c, Western blot and quantification of pro-IL-1β and tubulin (n = 3). Blot shows three biological replicates for each condition and data are mean normalized to NS and tubulin loading control +/- S.E.M. of biological replicates. d-h, BMDMs from wild-type (WT) and ND6P25L mice that were NS or stimulated with LPS for 3 h, 6 h or 24 h LPS (n = 3). d, e, Il10 and TNFα expression assessed by RT–qPCR (n = 3). f-h, IL-10, TNFα and IL-6 release measured by ELISA (n = 3). Data are mean +/- S.E.M. of biological replicates. P values displayed above graphs calculated using two-tailed Student’s t-test for paired comparisons.

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