Figure 5.
The anti-inflammatory activity of FCNG in THP1-MoM cells is mediated by downregulation of pro-inflammatory cytokines. (A–C) Gene expression analysis of IL-1β (A), IL-6 (B) and NFkB (C) by qPCR of THP1-MoM cells pre-treated with (11.7 ± 4.5) × 109 particles/mL of FCNP and FCNG for 24 h, followed by stimulation with LPS (100 ng/mL) for an additional 24 h. Non-stimulated cells were used as controls for LPS stimulation. (D,E) Total protein extracts of THP1-MoM cells treated as described in (A–C) were analyzed by Western blot to detect NFkB. The positions of relevant molecular mass markers (kDa) are indicated on the right side and GAPDH was used as the loading control. (E) Quantification of NFkB levels was performed by densitometry using ImageJ software, and is presented relatively to the GAPDH loading control as arbitrary units. Data are representative of three independent experiments and presented as mean ± SD. Two-way ANOVA and multiple comparisons were performed with the Dunnett’s test, and are presented relative to the LPS-stimulated cells. Statistical significance was defined as p ≤ 0.05 (*), p ≤ 0.01 (**), p ≤ 0.001 (***) and p ≤ 0.0001 (****).