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. 2018 Nov 27;9:1378. doi: 10.3389/fphar.2018.01378

FIGURE 5.

FIGURE 5

The RIPK1/toll-like receptor 4 (TLR4) pathway attenuates chondrocyte degeneration induced by Nec-1. (A,B) The expression levels of MMPs, ADAMTs5, and stromal cell-derived factor-1 (SDF1) in primary chondrocytes from the ctl, IL-1β, IL-1β + Nec-1, IL-1β + HMGB1, and IL-1β + HMGB1 + Nec-1 groups were determined by western blotting. (C) Serum levels of SDF-1α protein were measured by ELISA. Serum were collected from mice euthanized at 8 weeks post-DMM surgery (n = 10 for each group). (D,E) Interactions between HMGB1 and TLR4 were detected by co-immunoprecipitation. Relative protein expression was quantified using ImageJ software. The relative protein expression levels of HMGB1 and TLR4 were normalized to GAPDH and expressed as a relative value. (F,G) Expression of HMGB1 and TLR4 were determined by immunohistochemical staining of the cartilage from each group (n = 10). HMGB1 and TLR4 positive cells were quantified using Image-J software. The experiments were repeated three times independently. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.0006, and ∗∗∗∗P < 0.0001.