Pretreatment of D10 cells with AG-490 inhibits IL-4–induced STAT6 DNA binding as demonstrated by electrophoretic mobility shift analysis.

D10 cells were treated with the DMSO control (lanes A-D) or 100 μmol/L AG-490 (lanes E-H) and incubated with medium (−) or 100 nmol/L IL-4 (+) for 10 minutes at 37°C. Nuclear extracts corresponding to 5 μg of protein were incubated in the absence of antibody (lanes A, B, E, and F), α-STAT6 (lanes C and G), or normal rabbit serum (nrs; lanes D and H) and then with a phosphorus 32 (³²P)–labeled oligonucleotide probe corresponding to the Cε gene promoter. Arrow indicates migrational location of each nonsupershifted STAT6-DNA complex or free probe.