Figure 5. The effect of the systemic Akt1 or Akt2 deletion on neutrophil accumulation in the lungs of tumor-bearing mice.

A. Percentage of neutrophils in the lungs of nontumor-bearing mice after tamoxifen injection to systemically delete Akt1 or Akt2. The percentage of neutrophils was calculated by counting the number of Ly6G-positive cells relative to total hematoxylin and eosin-stained cells in lung tissue at endpoint sections as described in the Methods section. Data are presented as the means +/− SEM. P= 0.3, using unpaired t test. B. Quantification of lung neutrophils in control MMTV-PyMT mice and after systemic deletion of Akt1 or Akt2 at endpoint. Left: Representative lung section images stained with anti-Ly6G. Right: Quantification was performed after the primary tumors reached the endpoint. Data are presented as the means ± SEM. P=0.0066, MMTV-PyMT;Akt1^f/f;R26Cre^ERT2 vs. MMTV-PyMT;R26Cre^ERT2 and p=0.518, MMTV-PyMT;Akt2^f/f;R26Cre^ERT2 vs. MMTV-PyMT;R26Cre^ERT2 using an unpaired t test. Quantification of metastatic nodules (C) and neutrophils (D) in the lungs of R26Cre^ERT2 and Akt1^f/f;R26Cre^ERT2 mice, which were orthotopically transplanted with MMTV-PyMT (WT) tumor cells and injected with tamoxifen at palpation. Upper panel shows schematic of experimental design. Quantification was performed when the primary tumors reached the endpoint. Data are presented as the means ±SEM. P=0.019 for metastatic nodules, and P=0.0034 for neutrophils using an unpaired t test. Quantification of metastatic nodules (E) and neutrophils (F) in the lungs of R26Cre^ERT2 and Akt2^f/f;R26Cre^ERT2 mice, which were orthotopically transplanted with MMTV-PyMT (WT) tumor cells and injected with tamoxifen at palpation. Upper panel shows schematic of experimental design. Quantification was performed when the primary tumors reached the endpoint. Data are presented as the means ±SEM. P>0.05 for metastatic nodules and neutrophils using an unpaired t test.