Response to rapamycin of experimental tumors. (A) Shown on the vertical axis is the tumor volume in mm³ and on the horizontal axis the time since inoculation with 10⁶ BC-1 cells and simultaneous treatment with rapamycin (n = 5) or vehicle (n = 4). (B) Shown on the vertical axis is the tumor volume in mm³ and on the horizontal axis the time since inoculation with 10⁶ BC-1 cells and treatment with rapamycin (n = 5) or vehicle (n = 4) animals after the establishment of tumors (day 14). Error bars indicate the SD for each group of animal. For rapamycin-treated animals the error bar is smaller than the symbol. (C) Expression of IL-6 in a mock-treated (−) BC-1 tumor (red color), which is reduced by rapamycin treatment (+). Expression of IL-10 in a mock-treated BC-1 tumor (−) (red color), which is reduced by rapamycin treatment (+). No staining in the absence of the specific primary antibody (no 1°). All sections are counterstained with hematoxilin (blue) and are at 400 × magnification. (D) Representative immunohistochemistry of mouse xenograft tumors using antibodies specific for phospho-Akt (Akt-P), phospho-mTOR (mTOR-P), and phospho-p70S6 kinase (p70S6-P). The phospho Akt staining was performed on BCBL-1 tumors at either 100 × or 400 × magnification. No 1° indicates the no primary antibody control reaction. The phospho-mTOR and phospho-p70S6K staining was performed on 2 different tumors (BC-3a and BC-3b) derived from the BC-3 cell line; No 1° indicates the no primary antibody control reaction. Pictures here were taken at 400 × magnification. (E) Western lot analysis of protein extracts for the indicated cell lines exposed to rapamycin or vehicle using anti–phospho-S6 and anti-p70S6 kinase antibody. (F) Western blot analysis of protein extracts for the indicated cell lines exposed to rapamycin or vehicle using anti-mTOR and antiactin antibody.