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. 2018 May 23;14:150–157. doi: 10.1016/j.molmet.2018.05.014

Figure 2.

Figure 2

In vitro effects of GIP (1–42) on monocyte/macrophage inflammatory activation. Chemokine-induced migration of murine (RAW 264.7 cells) and human THP-1 monocytes. Monocytes were pretreated with GIP (1–42) for 30 min at the concentrations indicated before migration experiments using MCP-1 (10 nM) were performed in a modified Boyden chamber (A) (n = 3–4; r = 4), MMP-9 protein expression and activity analyzed by western blotting (IB) or zymography (Z) in the supernatant of RAW 264.7 cells 72 h after LPS stimulation (100 ng/mL) and 30 min pretreatment with GIP (1–42) at the indicated concentrations (B, C), IL-6 secretion of RAW 264.7 cells analyzed by ELISA 24 h after LPS stimulation (100 ng/mL) and 30 min pretreatment with 1 nM GIP (1–42) (D) (n = 4; r = 3), NF-κB p65 activation of RAW 264.7 cells at the indicated timepoints after LPS stimulation (100 ng/mL) and 30 min pretreatment with GIP (1–42) (n = 2; r = 3) (E) and western blot analysis of JNK, ERK, and p38 phosphorylation 30 min after LPS stimulation (100 ng/mL) and 30 min pretreatment with GIP (1–42) at the indicated concentrations (n = 3; r = 2). N: number of independent experiments; r: number of replicates. Results are expressed as the mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.001 LPS/MCP-1 vs. LPS/MCP-1 + GIP, #p < 0.05, ##p < 0.01 and ###p < 0.001 LPS/MCP-1 vs. control.