Research Paper Volume 11, Issue 23 pp 11659—11672

Nickel induces inflammatory activation via NF-κB, MAPKs, IRF3 and NLRP3 inflammasome signaling pathways in macrophages

Figure 5. NiCl2 activates NLRP3 inflammasome pathway in BMDMs. (A and B) Relative mtROS amounts determined by MitoSOX-red staining of NiCl2-primed BMDMs. Scale bar 50 μm. (C) Relative cytosolic mtDNA expression in NiCl2-primed BMDMs. (D) NiCl2-induced changes in mitochondrial membrane potential (Ψm) in BMDMs measured by TMRM fluorescence. (E) Immunoblot analysis of pro-caspase-1, cleaved-caspase-1, pro-IL-1β, cleaved- IL-1β, NLRP3 and ASC in lysates of NiCl2-treated BMDMs, and cleaved-caspase-1and cleaved- IL-1β in the supernatant. (F) Immunoblot analysis of pro-caspase-1, cleaved-caspase-1, pro-IL-1β, cleaved- IL-1β, NLRP3 and ASC in lysates of Mito-TEMPO (500 μM)-pre-treated 1h before 24h of NiCl2 stimulation. (G) Relative cytosolic mtDNA expression in NiCl2-treated (24h) BMDMs in the presence/absence of Mito-TEMPO (500 μM, 1h) pre-treatment. (H) Changes of mitochondrial membrane potential (Ψm) in NiCl2-treated (24h) BMDMs in the presence/absence of Mito-TEMPO (500 μM, 1h) pre-treatment. Data are presented with the means ± standard deviation (n=5). *p < 0.05 and **p < 0.01, compared with the control group.