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. 2015 Jan 21;6(6):4345–4356. doi: 10.18632/oncotarget.2999

Figure 4. PAK5-mediated GATA1 phosphorylation regulates EMT in breast cancer cells.

Figure 4

(A) MCF-7 cells were transfected with wild type or different mutants of GATA1 for Luciferase assay. **p < 0.01. (B) Empty vector, Flag-GATA1 wt or Flag-GATA1 S161A S187A was stably expressed in MCF-7 and NMuMG cells through lentivirus. Cell lysates from these cells were used for immunoblotting using anti-Fibronectin, E-cadherin, vimentin, GAPDH antibodies. (C) Stably knocked down shPAK5 or NC (non-specific control) cells were transfected with GATA1 wt. Western blot analyzed the marker of EMT. (D) The same transfection as (B), NMuMG cells were cultured 24 hours and analyzed by immunofluorescence using anti-E-cadherin antibody followed by Alexa Flour 488 (green) antibody and nucleus was stained by DAPI (blue). (E–F) Effects of different types of GATA1 over-expression on the migration of cultured MCF-7 cells were examined by wound-healing (E) and transwell migration chambers (F) assays. Results are representative of three independent experiments. Migrated cells were plotted as the average number of cells per field of view. In the low lane, transwell migration chambers were treated with 10% matrigel, but not in the top lane. *p < 0.05.