FIGURE 5.

IL-1β-mediated NP cells activated “M1” polarized THP-1 macrophages. (A).After being treated with IL-1β (20 ng/ml), NP cells were treated with a high concentration (100 μg/ml) of MAG. Then, THP-1 cells were treated with the CM of NP cells (CM^Control, CM^IL−1β, CM^{IL−1β+MAG(100 μg/ml)}) for 24 h. (B). QRT-PCR was implemented to verify the expression of inflammatory cytokines (including IL-1β, IL-6, TNF-α, and IL-18) in THP-1 cells. (C). ELISA was employed to determine the HMGB1 profile in the in the supernatant of THP1 cells. (D-F). The expression of MyD88-NF-κB and NLRP3-ASC-Caspase1 inflammasome was measured by WB. (G-H). Cell immunofluorescence was performed to detect p-NF-κB and NLRP3 in the TBP1 cells (×200). n = 3. ANOVA p < 0.01. **p < 0.01, ***p < 0.001 (vs CM^Control group). +p < 0.05, ++ p < 0.01 (vs CM^IL−1β group).