Figure 3.

Loss of GILZ increases the responsiveness of macrophages toward Pam₃CSK₄ and Poly(I:C). (A,B) Wildtype- (WT) and GILZ Knockout-(KO) BMMs were treated with Pam₃CSK₄ (4 h, A) or Poly(I:C) (6 h, B) at the indicated concentrations. TNF secretion was quantified by bioassay (n = 4, triplicates). (C–F) WT and GILZ KO BMMs were treated with Pam₃CSK₄ (100 ng/mL, C and E) or Poly(I:C) (10 μg/mL, D,F) for the indicated periods of time. ERK activation was determined by Western blot. (C,D) Representative blots. (E,F) pERK signal intensities were quantified and normalized to total ERK signals. Values for untreated WT cells (0′) were set as 1 (n ≥ 3). (G, H): WT and GILZ KO BMMs were pretreated with medium only (Med), the solvent control DMSO (0.1%), or U0126 (10 μM) for 2 h, followed by activation with Pam₃CSK₄ (100 ng/mL, 4 h; G) or Poly(I:C) (10 μg/mL, 6 h). TNF production was assessed by bioassay (Co: untreated cells; n = 4, triplicates). ^*p < 0.05, ^**p < 0.01, ^***p < 0.001.