Figure 1. Anti-metastatic potential of M2 in murine melanoma, B-16F10 cells in vitro and in vivo.

(A-B) Histopathological analysis of lung, liver and kidney in M2 treated and untreated B-16F₁₀ melanoma bearing C57BL/J mice. Representative H-E stained sections with arrows indicating metastatic colonies. (C) Effect of M2 on MMP activity analyzed by gelatin zymography. MMP-2 and MMP-9 activities were evidenced at the corresponding molecular weight. (D) densitometric analysis of MMP-2 and MMP-9 activities. (E-F) Wound-healing assay to assess the effect of M2 on cell B-16F₁₀ melanoma cell migration at three different time points (6, 12 and 24 hours) after M2 treatment. (G) Effect of M2 on invasion ability was measured using transwell assays in B-16F₁₀ melanoma cells. Microscopic observation (×10) of B-16 cells on the bottom of the Boyden's chamber at the end of the 24 h. (H) Optical Density (OD) and percentage of cells which have invaded to the lower chamber. (I) Effect of M2 on MMP activity analyzed by gelatin zymography. MMP-2 and MMP-9 activities were evidenced at the corresponding molecular weight. (J) Densitometric analysis of MMP-2 and MMP-9 activities. (K) qPCR data showing relative fold changes in gene expressions in M2 treated B-16 cells in comparison to untreated B-16 controls. (n=3 biological replicates, error bars denote s.e.m.). ^*p value<0.005, relative to Control.