1. Volume 21, Issue 9, Pages 2384-2392 (November 2017)
Hyperglycemia Impairs Neutrophil Mobilization Leading to Enhanced Metastatic Seeding 
Tanya Fainsod-Levi, Maya Gershkovitz, Sandra Völs, Saran Kumar, Saleh Khawaled, Jitka Y. Sagiv, Ronit V. Sionov, Myriam Grunewald, Eli Keshet, Zvi Granot 
Cell Reports 
Volume 21, Issue 9, Pages (November 2017)
DOI: /j.celrep
Copyright © 2017 The Author(s) Terms and Conditions

2. Cell Reports 2017 21, 2384-2392DOI: (10.1016/j.celrep.2017.11.010)
Copyright © 2017 The Author(s) Terms and Conditions

3. Figure 1 Enhanced Metastatic Burden in Hyperglycemic Mice
(A) Experimental timeline.
(B–D) Fed blood glucose levels (B), body weight (C), and tumor growth (D) in control, hyperglycemic, and normalized mice.
(E) Ki67+ cells in tumors from control, hyperglycemic, and normalized mice.
(F) CD31 immunohistochemistry in tumors from control, hyperglycemic, and normalized mice.
(G) Percent vascular coverage in tumors from control, hyperglycemic, and normalized mice.
(H and I) Average number (H) and representative images of lung metastases (I) in control and hyperglycemic tumor-bearing mice (day 28).
(J) Fraction of low-density neutrophils (LDN) in control and hyperglycemic tumor-bearing mice.
(K) Migration of neutrophils from control and hyperglycemic tumor-bearing mice.
(L) Extent of phagocytosis by neutrophil from control and hyperglycemic tumor-bearing mice.
(M) H2O2 production by neutrophils from control and hyperglycemic tumor-bearing mice in response to PMA.
(N) Cytotoxicity of neutrophils purified from control and hyperglycemic tumor-bearing mice.
∗p < 0.05, ∗∗p < Error bars represent SE.
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4. Figure 2 Metastatic Seeding Is Enhanced in Hyperglycemic Mice
(A) Average number of spontaneous metastases in control, hyperglycemic, and normalized mice (day 28).
(B) Average number of metastases following tail-vein injection of 4T1 cells in tumor-free control, hyperglycemic, and normalized mice (day 14).
(C and D) Average number of lung-associated GFP+ cells 48 hr (C) and 14 days (D) following tail-vein injection of GFP+-4T1 cells in control, hyperglycemic, and normalized tumor bearing mice.
(E) Average number of lung-associated GFP+ cells 48 hr after tail vein injection of GFP+-4T1 cells in tumor-free control, hyperglycemic, and normalized mice.
(F and G) Representative images of lungs from tumor-bearing control, hyperglycemic, and normalized mice 48 hr (F) and 14 days (G) after tail vein injection of GFP+-4T1 cells.
(H) Representative images of lungs from tumor-free control, hyperglycemic, and normalized mice 48 hr after tail vein injection of GFP+-4T1 cells.
∗p < 0.05, ∗∗p < Error bars represent SE.
Cell Reports  , DOI: ( /j.celrep )
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5. Figure 3
Decreased Numbers of Neutrophils in the Circulation and the Pre-metastatic Lungs of Hyperglycemic Mice
(A–D) Total number of circulating leukocytes (A), lymphocytes (B), monocytes (C), and neutrophils (D) in normoglycemic and hyperglycemic tumor-free and tumor-bearing mice (day 14).
(E and F) Representative images (E) and average length (F) of spleens from control, hyperglycemic, and normalized tumor-bearing mice (day 28).
(G and H) Average number (G) and representative immunohistochemistry images (H) of lung-associated Ly6G+ neutrophils in control, hyperglycemic, and normalized tumor-bearing mice (day 10).
(I and J) Average number (I) and representatvie immunohistochemistry images (J) of lung-associated Ly6G+ neutrophils in control, hyperglycemic, and normalized tumor-free mice.
(K–O) Total number of circulating leukocytes (K), CD4+ lymphocytes (L), CD8+ lymphocytes (M), monocytes (N), and neutrophils (O) in control, hyperglycemic, and normalized tumor-bearing mice (day 22).
(P) Circulating neutrophil numbers in normoglycemic and hyperglycemic mice treated with vehicle or rhG-CSF.
(Q and R) Circulating levels of G-CSF (Q) and GM-CSF (R) in normoglycemic and hyperglycemic tumor-bearing mice (day 10).
∗p < 0.05, ∗∗p < Error bars represent SE.
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6. Figure 4
Enhanced Metastatic Seeding in Hyperglycemic Mice Is due to Reduced Numbers of Neutrophils
(A) Experimental timeline for the 4T1 and Renca metastatic seeding assay.
(B) Flow cytometry analysis of Ly6G+ circulating neutrophils in control (immunoglobulin G [IgG]-treated) or neutrophil-depleted (anti-Ly6G-treated) normalized tumor-bearing mice.
(C) Representative immunohistochemistry images of lung-associated Ly6G+ neutrophils in control or neutrophil-depleted normalized tumor-bearing mice.
(D and E) Average number (D) and representative images (E) of lung-associated GFP+ cells 48 hr after tail vein injection of GFP+-4T1 cells in control, hyperglycemic, normalized (“Norm.”), and neutrophil-depleted normalized tumor-bearing mice (“Norm. Ly6G”).
(F and G) Average number (F) and representative images (G) of lung-associated GFP+ cells 48 hr after tail vein injection of GFP+-Renca cells in control, hyperglycemic, normalized, and neutrophil-depleted normalized tumor-bearing mice (“Norm. Ly6G”).
∗p < 0.05, ∗∗p < Error bars represent SE.
Cell Reports  , DOI: ( /j.celrep )
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