Evaluation of Stem Cell Signaling Modulators

Stem cell factor (SCF), stromal cell-derived factor 1 alpha (SDF-1α) and interleukin-3 (IL-3) were obtained from R&D systems (Minneapolis, MN). Chemical inhibitors PP3, EHT 1864, and Nexinhib20 were obtained from Tocris (Minneapolis, MN). Ascorbic acid and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) were obtained from Sigma-Aldrich (St. Louis, MO). MBQ-167 was obtained from Selleckchem (Houston, TX). Recombinant fibroblast growth factor 2 (FGF-2), antibody against β-Actin and the chemical inhibitors PP2 and GM6001 were obtained from EMD Millipore (Billerica, MA). Antibodies against Rab3B, Rab3D, Rab11A and Rab27A were obtained from Abcam (Cambridge, MA). Antibodies against Caveolin-1, Rab3A, Rab5 and Rab8 were obtained from Cell Signaling Technology (Danvers, MA). An antibody against RalA was obtained from BD Biosciences (San Jose, CA). GFP-RalA and S-Ch-Cdc42 adenoviruses were generated and utilized as described previously[13 (link), 40 (link)].