Figure 3.

CCL2 expression is regulated by constitutive Myd88-dependent signaling. (A) Myd88 was decreased in a stable cell line expressing shRNA specific for Myd88 (_consMyd^low) and assessed for Myd88 and CCL2 expression by qRT-PCR relative to a cell line expressing shRNA specific for lacZ as a control (cntrl). (B) Myd88 and CCL2 expressions were assessed by qRT-PCR in a cell line expressing shRNA specific for Myd88 under a tetracycline-inducible promoter upon no treatment (cntrl) or treatment with different doses of tetracycline (_indMyd^low) for 48 hours. (C) 4T1 cells were treated with 100 µM control peptide (cntrl) or Myd88-specific inhibitory peptide for 24–72 hours and examined for CCL2 expression by qRT-PCR. (D) 4T1 cells were treated with different doses of the control peptide (CP) and Myd88-specific inhibitory peptide (MP) for 72 hours and examined for CCL2 expression by qRT-PCR. Data were normalized to GAPDH and expressed relative to the control. All experiments were repeated at least three times, and where indicated (*), P < 0.05 using Student’s t-test relative to control.