Fig 1: Characteristics of G-MDSC cells and peritoneal cytokine after Sunitinib treatment. (A) Representative Wright-Giemsa staining of isolated peritoneal CD11b+ Ly6G+ G-MDSCs from at least 5 animals in each group after 7 days of treatment, Magnification 1000× Oil, and Scale bar 10µm. (B) Peritoneal cytokines VEGF, G-CSF, IL-6, TGF-ß after 7 days of treatment. Data are expressed as the mean ± SEM from 10 animals in each group, *P < 0.05.
Fig 2: Neutrophil accumulation involves in intestinal tumor deterioration in KRAS-mutated CRC; A, APC-WT and APC-KRASG12D mice at 8th, 12th, 16th and 20th weeks were euthanized. Tumor number was evaluated by counting single polyps in small colons. B, The number of neutrophils counted using flow cytometry, Single-cell suspensions were prepared from the peripheral blood (a), spleen (b), BM(c) and mLN (d) of APC-WT and APC-KRASG12D mice euthanized at indicated ages. The graphs show total numbers or frequencies of CD45.2 þ Ly6G þ CD11b þ neutrophils in respective organs. C, The content of G-CSF in the serum of APC-WT and APC-KRASG12D mice. * p < 0.05, n = 8. The measurement data was expressed as mean ± standard deviation. Data analysis at different time points was performed by repeated measurement ANOVA, follows by Bonferroni post hoc test
Fig 3: L971 ameliorates LPS induced septic shock in vivo. Mice were i.p. administrated with 5 mg/kg L971 12 h before LPS challenge (15 mg/kg, i.p.) and killed at 72 h after LPS injection. Survival rate (A) and body temperature (C) were recorded. In a separate experiment, mice were i.p. administrated with 2.5 mg/kg or 5 mg/kg L971 12 h before LPS challenge (15 mg/kg, i.p.) and killed at 36 h after LPS injection. Survival rate (B) and body temperature (D) were measured at 36 h after LPS treatment. Mice treated as in (A) were killed 72 h after LPS injection and monocyte (E) and neutrophil (F) counts in whole blood and plasma ALT levels (H) were measured. (G) Mice were i.p. administrated with 5 mg/kg L971 12 h before LPS challenge (15 mg/kg, i.p.) and killed at 6 h after LPS injection. The detection of plasma G-CSF level was evaluated by ELISA assays
Fig 4: C498 prevents temperature loss and tissue damage caused by LPS-induced septic shock in vivo. Mice were i.p. administrated with 5 mg/kg C498 12 h before the LPS challenge (6 mg/kg, i.p.). Body temperature (A) was recorded. Mice treated as in (A) were sacrificed 24 h after LPS injection and plasma ALT (B), AST (C), and BUN levels (D) were measured. Mice treated as in (A) were sacrificed 12 h after LPS injection and HE staining (100 ×) of inflammatory cell infiltration in the lung and kidney (E). Yellow arrows indicate neutrophil infiltration and black arrows indicate nuclei fragmentation. Mice treated as in (A) were sacrificed 12 h after LPS injection. Immunofluorescence (F, G) and relative fluorescence intensity (100 ×) of kidney and lung (H). The green color indicates CD11b staining and blue indicates nuclear DAPI staining (F-H). (I) The detection of 24 h plasma G-CSF levels by ELISA assay. *P < 0.05 is considered significant.
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