Fig. 4

Sirolimus treatment reduced p70S6K1 and 4E-BP1 phosphorylation A Representative mammary tissue section images (left) from paired pre- and post-treatment samples by patient number show immunohistochemically stained phosphorylated p70S6K1 (Thr389/Thr412) in normal ducts and DCIS lesions; nuclei are stained blue. The stained protein expression between paired tissue samples of pre- and post-treatment in normal (middle) (n = 8) and DCIS tissue (right) (n = 3) was scored in three representative high-power fields per tissue section as 0, no staining; 1 + , weak diffuse cytoplasmic staining, 2 + , moderate-to-strong granular cytoplasmic staining, and 3 + , cells stained with strong intensity. The Y-axis shows the % of each scored area. B Representative images of stained phosphorylated 4E-BP1 (Ser65/Thr70) and quantitative analysis of its expression in pre- and post-treatment normal (n = 9) and DCIS tissue (n = 5). An intensity score-weighted single value was calculated for each high-power field and used for comparison between pre- and post-treatment by 2-way ANOVA. Scale bar, 100 µm. Note: the fewer DCIS paired cases than normal duct paired cases were due to the absence of DCIS lesions in some tissue sections