FIGURE 2.

The K685R mutation impairs expression of U-STAT3-induced genes. A, hTERT-HME1 cells were infected with retroviral constructs expressing WT STAT3 or the STAT3 mutants K49R, K140R, K685R, or S727A. Stable pools of cells were selected with G418. Western analysis of total cell lysates was done to analyze STAT3 expression in these cells. B, the heat map of microarray performed to compare genes induced in hTERT-HME1 cells expressing wild-type STAT3 or STAT3 mutants shows the increased expression of 124 genes in response to increased levels of WT STAT3 and the changes in expression of these genes in cells with increased levels of each of the STAT3 mutants. In the heat map, red corresponds to high expression, green corresponds to low expression, and black corresponds to intermediate expression. We include only genes whose expression changed by >2-fold, with differential p ≤ 0.05, in WT cells when compared with vector control cells, and with average signals greater than 100 in WT cells. C, qPCR analysis of relative expression of CDC45L, IL7R, and CSF2 genes was done. Values are the means, with standard deviations, from triplicate experiments.