HMGB1 upregulated the expression of P-gp, TLR4, and RAGE in bEnd.3 cells.

<p>(A&B) The mRNA levels of <i>mdr1a</i> (A) and TLR4 (B) in bEnd.3 cells treated with different concentrations of HMGB1 were measured using q-PCR and normalized to the cells treated without HMGB1. HMGB1 at all tested concentrations (from 10 to 500 ng/mL) increased <i>mdr1a</i> mRNA and TLR4 mRNA levels in bEnd.3 cells as compared to cells treated without HMGB1. (C&D) The protein levels of P-gp (C), TLR4, and RAGE (D) in bEnd.3 cells treated with different concentrations of HMGB1 were analyzed using Western blotting with β-actin as a loading control. Protein levels were quantified and normalized to the cells treated without HMGB1. Treatment with HMGB1 at all tested concentrations increased P-gp, TLR4, and RAGE protein levels in bEnd.3 cells. Data were shown as mean±SD; <i>n</i> = 3. *<i>P</i><0.05, **<i>P</i><0.01 vs. cells treated with medium only. HMGB1, high-mobility group box-1; <i>mdr1a</i>, <i>multidrug resistance1a</i>; P-gp, P-glycoprotein; SD, standard deviation; TLR4, toll-like receptor 4; RAGE, receptor for advanced glycation end products.