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. 2016 Dec 12;213(13):2989–3005. doi: 10.1084/jem.20160817

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© 2016 Barrott et al.

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Figure 5. — Pten silencing associates with increased infiltration of myeloid-derived cells. (A) Example flow cytometry comparing an hSS;Pten^w/w tumor (top) to an hSS;Pten^fl/fl tumor (bottom) for F4/80⁺/GFP⁻ monocytes/macrophages (arrowhead, left plot), their MHCII⁺/Ly6C^high newly recruited monocyte (open arrow, middle plot) and MHCII⁺/Ly6C^mid tissue macrophage (filled arrowhead, middle plot) subpopulations, and Ly6C⁺/Ly6G⁺ neutrophils (arrowhead, right plot). (PE, phycoerythrin; APC, allophycocyanin; PerCP, perdinin chlorophyll protein). (B) Graph comparing the percent of infiltrating CD11B⁺ cells in hSS tumors of each Pten genotype (Student’s t test). (C) Representative low and high power photomicrographs and quantitation of CD68⁺ monocytes/macrophages (brown) by immunohistochemistry in hSS;Pten^w/w and hSS;Pten^fl/fl tumors (Student’s t test). (D) Representative low and high power photomicrographs and quantitation per high power field (HPF) of Leder cytochemically stained neutrophils (magenta) in hSS;Pten^w/w and hSS;Pten^fl/fl tumors (Student’s t test). (E) Representative photomicrographs of CD3 immunohistochemistry in hSS;Pten^w/w and hSS;Pten^fl/fl primary tumors and an associated graph demonstrating low and equivalent numbers of CD3⁺ lymphocytes (Student’s t test). (F) Pie charts presenting the mean fractions of neutrophils, monocytes, and lymphocytes in the peripheral blood of morbid tumor-bearing hSS;Pten^w/w and hSS;Pten^fl/fl mice (n = 7 and 13, respectively; Chi-squared test). (G) Graph of peripheral blood neutrophil counts in morbid tumor-bearing hSS;Pten^w/w and hSS;Pten^fl/fl mice (Student’s t test). Bars, 50 µm. Graph points present individual tumors; bars present means.