Figure 2
The IL-4 induction of mTARC is STAT6-dependent. A, Peritoneal thioglycollate-elicited Mθ from BALB/c wild-type or BALB/c STAT6-/- mice were treated +/- lOng/ml recombinant murine IL-4 for 24 hours. Total RNA was harvested and subjected to semi-quantitative RT-PCR with mTARC or β2M (housekeeping gene) primers. Chromatin immunoprecipitation experiments show STAT6 binding to the mTARC promoter. B, Bone marrow-derived Mθ from Balb/c and STAT6-/- were treated with 10 ng/ml recombinant murine IL-4 for the time periods shown before being formaldehyde fixed and sonicated. PCR using mTARC promoter primers was performed on DNA immunoprecipitated with STAT6 antibody in the presence or absence of a specific STAT6 blocking peptide. Amplification of 1/100th of input DNA and no input DNA controls were routinely performed in each ChIP experiment for positive and negative controls, respectively.