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. 2018 Mar 28;9:313. doi: 10.3389/fphys.2018.00313

Figure 4.

Figure 4

Functional role of HMGB1 receptors on the production of IL-1β in HMGB1-stimulated VSMCs. (A) VSMCs were transfected with siRNA (200 nM) for TLR2, TLR4, and RAGE, and then incubated with HMGB1 for 24 h. The levels of IL-1β in the culture media were quantified by ELISA. Data are expressed as means ± SEMs of triplicates pooled from 4 independent experiments. **P < 0.01 vs. non-treated control. ##P < 0.01 vs. corresponding value in negative control. (B) VSMCs were pretreated with anti-IgG antibody (10 μg/ml), anti-TLR2 antibody (10 μg/ml), anti-TLR4 antibody (10 μg/ml), or anti-RAGE antibody (10 μg/ml) for 30 min, and then stimulated with HMGB1 (100 ng/ml) for 24 h. IL-1β release into culture media was quantified by ELISA. Data are expressed as means ± SEMs of triplicates pooled from 4 independent experiments. (C) Protein levels of active IL-1β were assessed by Western blot using β-actin as an internal control. Data are expressed as means ± SEMs of duplicates pooled from 4 independent experiments. **P < 0.01 vs. non-treated control. #P < 0.05 and ##P < 0.01 vs. control in HMGB1-treated cells.