FIG. 8.

Stat5 and GR nuclear translocation. (A) HC11 cells were treated for the indicated times with different combinations of lactogenic hormones (dexamethasone [D], insulin (I), and prolactin [P]). Control cells (−) were left in serum-free medium. Nuclear and cytoplasmic fractions were prepared, and 80 μg of each was subjected to SDS-8% PAGE and probed with Stat5a and GR antisera. (B) HC11 cells were treated for the indicated times with lactogenic hormones; 80 μg of whole-cell protein extracts was subjected to SDS-8% PAGE and incubated with Stat5a and Stat5b antisera. (C) Cellular fractionation was controlled by probing two independent preparations of each fraction with a Raf-1-specific antiserum. Prolactin, insulin, and dexamethasone were at concentrations of respectively, 5 μg/ml, 5 μg/ml, and 1 μM, respectively.