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. 2016 Aug 24;17(9):1394. doi: 10.3390/ijms17091394

Figure 5.

Figure 5

CREB-mediated IL-6 production is involved in LPS-induced VSMC migration. (A) Serum-starved VSMCs were stimulated with LPS for the indicated times; (B,C) VSMCs were pretreated with different amounts of polymyxin B (B); SB202190 or U0126 (C) for 30 min and then stimulated with LPS for 30 min. Cell lysates were subjected to Western blotting with antibodies for CREB, phospho-CREB, NF-κB p65, phospho-NF-κB p65 (Ser536) or β-actin; (D–F) VSMCs were transfected with the indicated siRNAs for 24 h and then serum depleted for 24 h; (D) Cells were incubated with LPS for 30 min. Cell lysates were immunoblotted with antibodies for CREB, phospho-CREB, phospho-p38, phospho-ERK or β-actin; (E) Cells were stimulated with LPS for 24 h and IL-6 level in culture supernatants and CREB level in cell lysates were measured by ELISA and western blot, respectively; (F) Quiesced VSMCs were incubated with LPS for 24 h and migration assays performed as in Figure 2A. p < 0.001 vs. LPS + scRNA. The experiments in A–D were repeated three times with similar results. Data in E–F represent mean ± SD of three experiments. Statistical analyses were performed using the one-way ANOVA.