The number of EPCs was expressed per 1 mlblood [22]. Figure 1 Characterization of endothelial progenitor cells (EPCs) by flowcytometry evaluation. First, cells were plotted in forward vs side scatter to gate the lymphocyte population selectively, where EPCs are usually found (a). For analysis of CD45dimCD34+KDR+ endothelial progenitor cells, CD45 was then plotted against the side scatter (b), followed by further analysis learn more of the CD45dim population on coexpression of CD34/KDR (c). Nitrite and leptin measurement Mice were

fasted for 14 h prior to sacrificing in order to obtain fasted blood samples. Plasma was isolated from whole blood collected and total nitrite (NOx) was measured (R&D Systems) as an indicator of endothelial release of NO as previously described [23]. Moreover, plasma leptin concentration was measured by ELISA kit (R&D Systems) in mice according to manufacturer’s instructions. Statistical analysis Data are expressed as mean ± SD and were tested for normal distribution with the Kolmogorov-Smirnov test.

Comparisons between groups were analysed by ANOVA followed by the Bonferroni method as post hoc-test. Differences in the weight of the mice were analyzed using the paired-sample t test. Statistical significance was assumed, if a null hypothesis could be ARS-1620 cell line rejected at p ≤ 0.05. All statistical analysis was performed with SPSS 16 (SPSS Inc.). Results The plasma Lazertinib nmr levels of leptin were significantly higher in leptin group compared to all other groups of mice while there was

no significant difference between other groups (Figure 2). Figure 2 The plasma levels of leptin were significantly higher in leptin group compared P-type ATPase to all other groups of mice while there was no significant difference between other groups. * (p < 0.05). Body weights for each group of mice are shown in Table 1. There was a significant weight loss in mice of leptin group while the weight of the animals of 9F8 group increased significantly during the study. By the end of the experiment there was a significant difference between leptin and 9f8 group in body weight and also between each group and its relevant control group. Table 1 The weight of mice in each group of the study. group Mice weight1 Mice weight2 P(before-after) IgG 23.41 ± 0.31 23.24 ± 0.479 p > 0.05 9f8 22.74 ± 0.30 25.37 ± 0.77* P < 0.05 leptin 22.68 ± 0.99 19.25 ± 1.53*γ P < 0.05 PBS 24.37 ± 1.22 24.60 ± 1.20 p > 0.05 *Significant difference with respective control group γ Significant difference with 9F8 group The melanoma tumor weight of leptin treated mice were significantly more than tumors from other groups of mice while there was no significant difference between other groups (Figure 3). Figure 3 Mean tumors size and weight. The weights and volume of melanoma tumors excised from leptin treated mice were significantly larger than tumors from other groups of mice. There was no significant difference between three other study groups. * (p < 0.05).