Culture of Drosophila S2 cells and their use for RNAi-mediated loss-of-function studies and immunofluorescence microscopy.
Cultured Drosophila cell lines have become an increasingly popular model system for cell biological and functional genomic studies. One of the most commonly used lines, S2 cells, is particularly useful as it is easy to grow and maintain in the lab, is highly susceptible to gene inhibition using RNAi and is well suited to high-resolution light microscopic assays. Here, we provide protocols for the routine culture and RNAi treatment of S2 cells and methods to prepare these cells for fluorescence microscopy. Using these techniques, loss-of-function experiments may be performed after 4-7 d of RNAi-mediated protein depletion.
Rogers
Stephen L
SL
Department of Biology, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27515, USA. srogers@bio.unc.edu
Rogers
Gregory C
GC
D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D004331:Drosophila melanogaster / Q000166:cytology*; D008856:Microscopy, Fluorescence / Q000379:methods*; D034622:RNA Interference