Skip to main content
. 2020 Oct 9;8(10):403. doi: 10.3390/biomedicines8100403

Figure 4.

Figure 4

Stearic acid cooperative effect with TNF-α for MIP-1α/CCL3 production requires IRF3. (A) THP-1 monocytic cells were transfected with either control or IRF3 siRNA and incubated for 36 h. Real time PCR was done to measure IRF3 expression. (B,C) IRF3 deficient THP-1 cells were stimulated with stearic acid and TNF-α. MIP-1α/CCL3 expression was determined. The results obtained from minimum three independent experiments with three replicates of each experiment are shown. (D) IRF3 activity reporter monocytic cells were treated with stearic acid (200 µM) or 0.1% BSA (control) or TNF-α (10 ng/mL) or in combination. Culture media were collected after 24 h. Cell culture media were assayed for luciferase activity representing the degree of IRF3/ISRE activation using Quanti-Luc medium. (E) Western blot analysis showed that stearic acid induced IRF3 phosphorylation in a time dependent manner in THP-1 monocytes, verifies the role of IRF3 in the cooperative effect of stearic acid in the TNF-α mediated production of MIP-1α/CCL3. (F) Expression of phosphorylated IRF3 is shown as determined by densitometry of western blot bands. * P < 0.05; ** P < 0.01; *** P < 0.001.