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. 2015 Dec 23;6(42):44927–44940. doi: 10.18632/oncotarget.6743

Figure 3. TMSB4Y expression in Dox-inducible clones.

Figure 3

The TMSB4Y cDNA was cloned into the pBI-EGFP vector and stably transfected into the Dox-inducible MCF-10A cell line, TetHyg2.5, to the create TMSB4Y Dox-inducible clones TmY1 and TmY2. HEK293 cells were transiently transfected with empty vector (293 Empty Vector) or a TMSB4Y cDNA (293 TMSB4Y) to serve as negative and positive antibody controls, respectively. Parental TetHyg2.5, a stably transfected empty vector (EV) control cell line and the two inducible clones, TmY1 and TmY2, were placed in un-induced (−Dox) and induced (+Dox) media conditions for 48 hours prior to harvesting for lysates and fixation for FFPE blocks. TMSB4Y protein expression was verified via A.) western blotting using an anti-TMSB4Y antibody and GAPDH antibody as loading control, and B.) immunohistochemistry labeling of FFPE cell pellets using an anti-TMSB4Y antibody and counterstaining. Original magnification: 20X.