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. 2015 Oct 20;10(10):e0140918. doi: 10.1371/journal.pone.0140918

Fig 4. HMGB1 upregulated the expression of P-gp, TLR4, and RAGE in bEnd.3 cells.

Fig 4

(A&B) The mRNA levels of mdr1a (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 mdr1a 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; n = 3. *P<0.05, **P<0.01 vs. cells treated with medium only. HMGB1, high-mobility group box-1; mdr1a, multidrug resistance1a; P-gp, P-glycoprotein; SD, standard deviation; TLR4, toll-like receptor 4; RAGE, receptor for advanced glycation end products.