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

FIGURE 3.

FIGURE 3.

Ser-727 of MLS-Stat3 affects colony formation in soft agar, cell invasion, and tumor growth in mice. A, wild type Stat3 or Stat3 containing an MLS with mutations in the nuclear localization sequence, DBD, SH2, and Tyr-705 (mutMLS-Stat3) were expressed in 4T1 cells and assayed for their ability to form colonies in soft agar. mutMLS-Stat3 cells with Ser-727 mutated to alanine formed fewer colonies than cells with Ser-727 not mutated or mutated to aspartic acid. *, p < 0.05 versus Ser-727; #, p < 0.05 versus S727A (one-way ANOVA). B, Matrigel invasion assays were performed using 4T1 cells expressing various forms of Stat3. Single mutations in Y705F (YF), the DNA binding domain, and the SH2 domain (SH2) are the green bars. *, p < 0.05 versus MSCV; #, p < 0.05 versus S727A (SA); &, p < 0.05 versus S.D. (one-way ANOVA). C, tumor volumes at the site of injection of BALB/c mice with cell lines expressing various forms of MLS-Stat3. Cell lines that express Stat3 with an MLS in addition to the indicated mutations were injected into mice. Volumes were calculated using calipers. The animals were sacrificed after 27 days. *, p < 0.05 versus MSCV; #, p < 0.05 versus SA; &, p < 0.05 versus S.D. (one-way ANOVA). D and E, numbers of metastatic colonies in the livers and lungs formed in mice injected with cells expressing various forms of Stat3. *, p < 0.05 versus MSCV; #, p < 0.05 versus S727A (SA); &, p < 0.05 versus S.D. (one-way ANOVA).