Figure 3. New protein synthesis is required for inhibition of CIITA and CXCL11.

BMDM were pretreated with 500 nM cycloheximide or DMSO for 1 h prior to treatment with 10 ng/ml Pam₃CSK₄ for 8 h followed by 20 ng/ml IFN-γ for 4 h in the continued presence or absence of inhibitor. Total RNA was harvested after IFN-γ stimulation. CXCL11 (A), CIITA (B), and NOS2 (C) expression was assayed by qPCR and normalized to GAPDH and untreated samples. The concentration of cycloheximide used inhibited TNF production (as a measure of protein synthesis) by over 90% with minimal cell death. Statistical significance between IFN-γ alone samples and those treated with Pam₃CSK₄ prior to IFNγ was determined by two-tailed t-test.