Fig. 1.

DGLA attenuates IFN-γ induced MCP-1 and ICAM-1 expression in human and mouse macrophages.

THP-1 macrophages (A–B), primary HMDM (C–D) or mouse RAW264.7 macrophages (E–F) were pre-incubated for 24 h with the indicated concentrations of DGLA (A–B) or 50 μM DGLA (+, C–F) or DMSO vehicle (−). The cells were then incubated for 3 h with 250 U/ml IFN-γ (+) or its vehicle (−). Total RNA was subjected to RT-qPCR using primers against MCP-1 (A, C, E), ICAM-1 (B, D, F) or the housekeeping genes [human GAPDH (A–D) or mouse β-actin (E–F)]. The mRNA levels were calculated using the comparative Ct method and normalized to the housekeeping gene with values from cells pre-incubated with DMSO vehicle and then treated with IFN-γ given an arbitrary value of 1. Graphs display normalized gene expression (mean ± SEM) from three independent experiments. Statistical analysis was performed using a One-way ANOVA (equal variances) with Tukey's post hoc analysis (*, p ≤ 0.05; ***, p ≤ 0.001).