Fig. 3

BRAF oncogenic pathway activation selectively up-regulates NAMPT expression in MM. A, B BRAF WT MM cell lines MEL-505 (A) and Mewo (B) were infected with a lentivirus carrying the BRAF V600E construct. A control (CTRL) vector was used as comparison. Panels on the left show representative western blot analysis of BRAF V600E, phosphorylated (p)ERK1/2 and NAMPT expression. Actin was used as loading control. In the middle of the Figure NAMPT expression (green fluorescence), evaluated by confocal staining (original magnification ×63, scale bar: 25 µm). Histogram plots represent cumulative data of NAMPT expression (intensity) measured from at least 3 different fields of 3 pictures of 3 independent confocal experiments comparing CTRL (white bar) vs. V600E cells (red bar). Statistical significance was calculated using Mann–Whitney test. Heatmaps on the right show cumulative data of qRT-PCR analysis of the expression of NBEs i.e., NAMPT/NAPRT/NMRK1/QPRT (mean of at least 5 independent experiments) in all cell line variants. Data in the Figure are presented as the mean ± SEM. Significance was represented as: *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001