Fig. 4

Runx2 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 6 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 6 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 cells