N-WASp Knockdown MTLn3 Cell Line Expressing Dendra2

Growth Conditions: Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow VIII (TM018) + 5% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. Selection with 1 mg/ml G418 (G271) and 10 µg/ml Puromycin (G264)

Lipofectamine2000 transfection of Dendra2 plasmid (pDendra2-N ) with photoconvertible fluorescent marker in MTLn3 cells followed by selection for stable Dendra2 expression using 1 mg/ml geneticin. Lipofectamine2000 transfection of the pSUPERretro N-WASp plasmid into MTLn3 Dendra2 cells followed by selection for stable expression of N-WASp shRNA using 10 µg/ml puromycin.