Skip to main content
. 2018 Aug 13;9:1871. doi: 10.3389/fimmu.2018.01871

Figure 8.

Figure 8

Knockdown of Cdc42 Rho GTPase suppresses LL-37-induced chemokine production, but not anti-inflammatory cytokine IL-1RA. Human monocytic THP-1 cells were treated with either human Cdc42 Accell SiRNA smartpool (1 µM) or non-target control (NTC) Accell SiRNA smartpool (1 µM) for 96 h. The efficiency of knockdown was determined by western blots probing with Cdc42 antibody and protein loading using actin antibody, after (A) 96 h of SiRNA delivery, and subsequently following cell differentiation to plastic-adherent, macrophage-like THP-1 cells after additional (B) 24 and (C) 48 h. Macrophage-like THP-1 cells, either wild type, knockdown (KD), or cells treated with 50 µl SiRNA buffer (BC), were stimulated with LL-37 (5 µM), sLL-37 (5 µM each) or lipopolysaccharide (10 ng/ml). Tissue culture supernatants were monitored by ELISA for the production of chemokines (D) GRO-α and (E) IL-8 after 24 h, and for (F) anti-inflammatory cytokine IL-1RA after 48 h. Results shown as mean ± SE of three independent experiments (n = 3). Analysis of variance with Bonferroni’s post hoc test was used for statistical analyses (**p < 0.005, ***p < 0.0005).