Fig. 4From: USP24-dependent stabilization of Runx2 recruits a p300/NCOA3 complex to transactivate ADAMTS genes and promote degeneration of intervertebral disc in chronic inflammation miceRunx2 was phosphorylated in LPS-IVDs and LPS-treated NP-1/AF-1 cells(A) Protein levels of NCOA3-p300-Runx2 members in IVDs from sham- and LPS-treated mice. Homogenates of three lumbar discs (L1/L2) from three sham- and LPS-treated mice were used for western blotting to detect the protein levels of NCOA3, p300, Runx2, and GAPDH (loading control). (B and C) Protein levels of NCOA3-p300-Runx2 members in LPS-treated NP and AF cells. Three NP/AF cell lines (1, 2, and 3) were incubated with or without 20 ng/mL LPS for 6Â h. Cell lysates were used for western blotting to determine the protein levels of NCOA3, p300, Runx2, and GAPDH (loading control). (B) NP cells; (C) AF cells. (D and E) Protein levels of NCOA3-p300-Runx2 members in NP and AF cells co-treated with LPS and phosphatase. Three NP/AF cell lines (1, 2, and 3) were incubated with or without 20 ng/mL LPS and 200 units of phosphatase for 6Â h. Cell lysates were used for western blotting to determine the protein levels of NCOA3, p300, Runx2, and GAPDH (loading control). (D) NP cells; (E) AF cells. (F-H) Protein levels of different kinases in LPS-treated IVDs and LPS-treated NP-1/AF-1 cells. The same protein samples as in (A-C) were used for western blotting to determine the protein levels of p38, ERK1, ERK2, JNK1, and GAPDH (loading control). (F) IVDs; (G) NP cells; (H) AF cellsBack to article page