Skip to main content
. 2010 Aug 31;5(8):e12265. doi: 10.1371/journal.pone.0012265

Figure 1. EMMPRIN up-regulates VEGFR-2 expression in primary melanoma cells.

Figure 1

A. M10 and WM278 cells were transfected for 24 hours with two different EMMPRIN siRNA (Ambion siRNA) or their corresponding scrambled control siRNA (Ctl siRNA) at 33 nmol/L concentration. EMMPRIN and VEGFR-2 protein expression was evaluated by Western blot analysis of 15 µg cell lysates respectively (actin was used as a loading control; representative blot of 3 independent experiments is shown) and conditioned medium (CM) was harvested for gelatinases and uPA zymographies. B. Immunofluorescence of M10 cells treated with EMMPRIN siRNA and immunostained for EMMPRIN, VEGFR-2, uPA and TIMP-1. C. QRT-PCR analyses of EMMPRIN, VEGFR-2 and TIMP-1 showing the mean ±SD of relative expression to PPIA house keeping gene of at least 3 independent experiments. D. M10 and WM278 cells were incubated with CHO-membranes containing or not EMMPRIN (Ctl and Emp respectively). VEGFR-2 and uPA transcript levels were quantified by qRT-PCR showing the mean ±SD of relative expression to PPIA house keeping gene of at least 3 independent experiments. VEGFR-2 protein expression was evaluated by Western blot analysis of 30 µg cell lysates (actin was used as a loading control; representative blot of 3 independent experiments is shown); bars, SD. *, p<0.05.