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. Author manuscript; available in PMC: 2013 Jun 1.
Published in final edited form as: J Immunol. 2012 Apr 20;188(11):5682–5693. doi: 10.4049/jimmunol.1103760

Figure 6. C1q decreased procaspase-1 and pro-IL-1β cleavage in LPS-stimulated HMDMs.

Figure 6

HMDMs were incubated with PKH26-labeled EAL and LAL, pre-incubated with C1q, at a 5:1 ratio for 1 h and then stimulated with 10 ng/ml LPS. (A–B) HMDMs were stimulated with LPS for 6 h. ATP (1 mM) was added 90 min before the end of the stimulation. Cleaved caspase-1 was detected by FITC fluorescent caspase-1 probes and HMDMs were stained with a blue cell tracker. Representative merged micrographs of 3 independent experiments (from 3 different donors) are shown. Scale bar = 50 µm. Areas in white boxes were enlarged to show PKH26-AL (top, red), cleaved caspase-1 (middle, green) and the merge (bottom). (B) Quantification of cleaved caspase-1 in HMDMs. (C) NLRP3, procaspase-1, ASC and actin expression in HMDM cell extracts. Representative blots of 2 independent experiments are shown. (D–E) Levels of pro-IL-1β relative to β-actin levels (D, cell extracts) and mIL-1β relative to pro-IL-1β (E, supernatants) in HMDMs stimulated with LPS for 18 h with ATP added during the last 3 h of stimulation. Representative blots of 3 independent experiments are shown. All results represent means ± s.d. (n = 3 different donors), two-way ANOVA, * p < 0.05, ** p < 0.01 and ***, p < 0.001.