Fig. 5.

Liver kinase B1 (LKB1) inhibits high phosphate-induced calcification via HMGB1. Vascular smooth muscle cells (VSMCs) were infected with adenovirus-expressing green fluorescent protein (GFP), LKB1, and/or HMGB1, then treated with high phosphate for 6 days. A. Protein levels of Runx2, LKB1, and HMGB1, as detected by Western blot analysis (n = 3). B. Representative Alizarin Red staining in VSMCs (n = 3). C. Total calcium content in VSMCs (n = 5). D. Alkaline phosphatase (ALP) activity in VSMCs (n = 6/GFP and LKB1 groups, n = 3/HMGB1 and LKB1+HMGB1 groups). E–K. mRNA levels of LKB1 (E), HMGB1 (F), alkaline phosphatase ligand (ALPL) (G), Runx2 (H), Msx2 (I), Wnt3a (J), and Wnt7a (K), as detected by RT-PCR (n = 4). *P < 0.05, **P < 0.01, ***P < 0.001 vs. GFP; #P < 0.05, ##P < 0.01, ###P < 0.01 vs. HMGB1. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)