Figure 4

MIF knockdown decreases anchorage-independent colony formation. Three days after transfection using siRNA duplexes, melanoma cells were harvested and seeded into soft agar and permitted to form colonies according to the Methods. Representative low power photomicrographs demonstrating the relative abundance of colonies formed by (A) MelCV and (B) Me1007 cells after control NC or MIF siRNA knockdown (the insets show colonies in detail under higher magnification). The frequency and size of colonies were then estimated for MelCV (C) (E), respectively and Me1007 (D) (F), respectively where there was a significant reduction in colony numbers and size after MIF knockdown. Values are mean +/-S.E.M. (n = 3, ****p < 0.0001 compared to siNC transfected cells using Student’s t-test).