Fig. 3. TNF-α mediated pro-inflammatory phenotypic shift requires ACSL1. (A) Monocytes were transfected with Neg-siRNA or siACSL1 siRNA and incubated for 36 hours. Real-time PCR was performed to measure ACSL1 expression. ACSL1-deficient cells were treated with TNF-α and vehicle. (D, G, J) CD11c, CD11b, HLA-DR mRNA was determined by real time RT-PCR. Cells were stained with antibody against CD16, CD11c, CD11b, or HLA-DR, along with matched control antibody, and assessed by flow cytometry. (B, E, H, K). Flow cytometry data are presented as a bar graph of mean staining intensity (SI) as well as (C, F, I, L) representative histogram. Bar graphs depict mean values ± SEM of staining intensity (SI). P<0.05 was considered as statistically significant (* P<0.01; **P< 0.001, ***P< 0.0001). The data in all Fig.s are representative of three independent experiments.