Fig. 2

Aldh1a1 expression and activity can be downmodulated in B16F10 melanoma cells. Wild type B16F10 melanoma cells were retrovirally transduced with a sh-RNA molecule as indicated in the Methods to produce the sh-Aldh1a1 cell line. a Aldh1a1 mRNA levels were determined in wild type B16F10 and sh-Aldh1a1 cells by RT-qPCR. b Aldh1a1 activity was quantified by flow cytometry in both cells lines using the Aldefluor reagent kit. c B16F10 and sh-Aldh1a1 cells were analyzed for cell cycle distribution by flow cytometry. The percentage of cells in each cell cycle phase is indicated. d The kinetics of cell proliferation were determined by counting de number of cells during a 72 h period. e The migration of B16F10 and sh-Aldh1a1 cells was analyzed using wound healing assays. The B16F10 cell line was also transduced with a retrovirus containing the empty vectors used to produce the sh-Aldh1a1 cell line. Measurements were done in triplicate in two different cultures. At least 6 fields were analyzed for each wound in panel (e). Data are shown as mean ± SD. A.U. stands for arbitrary units