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. Author manuscript; available in PMC: 2017 Mar 1.
Published in final edited form as: Nat Immunol. 2016 Jun 27;17(9):1037–1045. doi: 10.1038/ni.3509

Figure 7.

Figure 7

Sensing of bacterial viability induces MyD88/TRIF- and NLRP3 inflammasome-dependent decrease in supercomplex abundance and increase in CII activity. (a) BN-PAGE immunoblot analysis and CI-in gel activity (IGA) assay in BMDMs treated as indicated. (b-e) SRC (b), ECAR (c) and CII activity (d, e) in BMDMs (b-d) and CD14+CD16- human monocytes (e) stimulated with EC or HKEC are shown. (f) ROS production by BMDMs stimulated as in (b). (g, h) CII activity in WT BMDMs stimulated as indicated for 1.5h. (i) Substrate-driven ATP synthesis assay in WT BMDMs stimulated with pI:C. 100% activity corresponds to a rate of 25.1 and 19.8 nmol ATP/min/mg protein for CI- and CII-mediated synthesis. (j) CII activity in WT, Trif-/-, Myd88-/- and Trif-/-Myd88-/- BMDMs treated as indicated. (k) BN-PAGE immunoblot analysis of resting and E. coli-stimulated WT, Trif-/- and Myd88-/- BMDMs. Arrow indicate the main SCs affected. (l, n) Densitometric analysis of CI+CIII2/CII signal ratio as observed by BNGE immunoblot of BMDMs of the indicated genotype stimulated with EC for 1.5h. (m) CII activity in WT, Nlrp3-/- and Caspase1-/- Caspase11-/- BMDMs stimulated with EC for the indicated time point. NS, not significant; *P < 0.05; **P < 0.01; ***P < 0.001 (two-tailed unpaired Student’s t-test). Data (mean and s.e.m. (b-j, l-n)) are from three (b, c, e, i), four (d, g, h, m, n) or five (f, j, l) independent experiments performed in two (d, e, g, h, j), three (f, i) or five (b, c) technical replicates, one representative of four independent experiments (a, k).