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. 2013 Sep 9;288(43):31280–31288. doi: 10.1074/jbc.M113.505057

FIGURE 2.

FIGURE 2.

A, expression of various forms of MLS-Stat3 in 4T1 cells does not alter OSM-stimulated accumulation of C/EBPδ mRNA. 4T1 cells were incubated with OSM for 4 h. RNA was collected and analyzed for the expression of C/EBPδ by quantitative PCR. Samples were normalized to the expression of actin mRNA. *, p < 0.05 versus corresponding control; two-tailed Student's t test. B, expression of Stat3 (Y705F) without an MLS in 4T1 cells inhibits OSM-stimulated accumulation of C/EBPδ RNA. 4T1 cells were incubated with OSM for 4 h. RNA was collected and analyzed for the expression of C/EBPδ by quantitative PCR. Samples were normalized to the expression of actin mRNA. *, p < 0.05 versus MSCV control (CTL) group (two-way ANOVA). C, tyrosine phosphorylation of endogenous Stat3 is altered by the expression of mutMLS-Stat3(S727D). Cytosol, mitochondria, and nuclear extracts were prepared from 4T1 cells expressing mutMLS-Stat3 with Ser-727, S727A, or S727D. Western blots were probed for tyrosine-phosphorylated Stat3 (upper panel), serine-phosphorylated Stat3 (second panel), and total Stat3 (third panel). The blots were also probed for tubulin (cytosolic marker), porin (mitochondrial marker), and lamin (nuclear marker).