Figure 1
Mutually exclusive expression of nuclear-localized pYStat5 and GLUT1 in human breast cancer. (A) Five human breast cancer cell lines were treated with prolactin or vehicle for 15 min at pHe 7.4 or pHe 6.8. Cell lysates were immunoprecipitated with anti-Stat5, resolved by SDS-PAGE and immunoblotted with anti-pYStat5 or anti-Stat5. (B) Median GLUT1 and Nuc-pYStat5 levels (AQUA scores) of the 52 breast cancer specimens in Cohort I are shown. Red dashed lines indicate the cutoff values for GLUT1 positivity and Nuc-pYStat5 positivity. (C) Example of a human invasive ductal carcinoma showing regional variability of pYStat5 IHC staining intensity. (D) GLUT1 and Nuc-pYStat5 AQUA scores of 2,244 randomly sampled tumor regions from 52 breast cancer specimens of Cohort 1. Examples of co-staining images show high GLUT1/low Nuc-pYStat5 (panel 1), high GLUT1/high Nuc-pYStat5 (panel 2), or low GLUT1/low Nuc-pYStat5 (Panel 3). (E) Cellular GLUT1 and Nuc-pYStat5 intensities in 8,804 cells sampled from six tumor spots displaying both high GLUT1 and high Nuc-pYStat5 are scatter-plotted. Representative images used for the analysis are shown. AQUA, automated quantitative analysis; GLUT1, glucose transporter 1; IHC, immunohistochemistry; Nuc-pYStat5, nuclear localized and tyrosine phosphorylated Stat5; pHe, extracellular pH.