Fig. 3. Characteristics of OPN expression in HMGB1-stimulated VSMCs.
(A) VSMCs were stimulated with HMGB1 (100 ng/ml) for 0 to 24 h, and then the expression of OPN mRNA was determined by RT-PCR. GAPDH was used as an internal control. The relative intensities of OPN mRNA to GAPDH were quantified and expressed as the means ± SEMs of 4 independent experiments. **p < 0.01 vs. value at 0 h. (B) In VSMCs treated with HMGB1 (100 ng/ml) for 0 to 48 h, the expression of OPN protein was determined using a Western blot assay. β-Actin was used as an internal control. The relative intensities of OPN protein to β-actin were quantified and expressed as the means ± SEMs of 4 independent experiments. *p < 0.05 and **p < 0.01 vs. value at 0 h. OPN, osteopontin; VSMCs, vascular smooth muscle cells; HMGB1, high-mobility group box 1; RT-PCR, reverse transcription polymerase chain reaction.
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