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. 2016 Aug 22;7(39):63651–63660. doi: 10.18632/oncotarget.11467

Figure 2. Up-regulation of TLR2-mediated expression of IL-23p19 by NOD2 in PC-like cells.

Figure 2

A. PC-like cells were stimulated for 4 h with MDP (10 μg/ml) only; B and C. PC-like cells were stimulated for 4 h with the TLR2 agonist PGN (10 μg/ml) (B) and Pam3CSK4 (1 μg/ml) (C) with or without MDP (10 μg/ml); D-F. PC-like cells transfected with transfection reagent only (Mock) or NOD2 siRNA (50 nM) were stimulated for 4 h with PGN (10 μg/ml) (D), Pam3CSK4 (1 μg/ml) plus MDP (10 μg/ml) (E) and Pam3CSK4 (1 μg/ml) (F); G. PC-like cells were stimulated for 4 h with Pam3CSK4 (1 μg/ml) or PC-like cells transfected with NOD2-siRNA were stimulated for 4 h with PGN (10 μg/ml); (A-G) total RNA was isolated and IL-23p19 mRNA expression was determined by real-time PCR and normalized to 18 S rRNA. H. PC-like cells transfected with transfection reagent only (Mock) or NOD2 siRNA (50 nM) were stimulated with PGN (10 μg/ml) for 4 h and finally whole-cell extracts were analyzed for IL-23p19 by western blot. Top, quantitative analysis of proteins; bottom, representative immunoblot images. I. Transfection efficiency was tested after 72-h transfection with siRNA by western blot. Ctrl represents untreated PC-like cells. Data are shown as means ± SD of three independent experiments, n.s., not significant; *P < 0.05; **P < 0.01; ***P < 0.001.