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. 2010 Oct 7;285(49):38093–38103. doi: 10.1074/jbc.M110.162388

FIGURE 5.

FIGURE 5.

STAP-2 PH domain regulates Brk-induced tyrosine phosphorylation of STAT3 at Tyr-705 and nuclear co-localization with Brk. A, 293T cells in a 12-well plate were transfected with FLAG-tagged Brk (1.0 μg) with or without Myc-tagged STAP-2 deletion mutants (1.0 μg). At 48 h after transfection, the cells were lysed and immunoblotted (IB) with an anti-pSTAT3 (Tyr-705), anti-STAT3, anti-PY (short exposure and long exposure for ECL detection), anti-FLAG, or anti-Myc antibody. B, HeLa cells in 12-well plates were transfected with FLAG-tagged Brk (1.0 μg), together with Myc-tagged STAP-2, or Myc-tagged STAP-2 ΔPH (1.0 μg) using jetPEI. At 48 h after transfection, the cells were fixed, incubated with anti-FLAG or anti-Myc antibodies, and visualized with FITC- and rhodamine-conjugated secondary antibodies. The same slides were also stained with DAPI to detect the nuclei.