GGA depletion in transgenic RNAi fly lines. (A) GGA was depleted ubiquitously in 2 fly lines [insertions on chromosome (2) or chromosome (3)] using a Tubulin-Gal4 driver, and whole fly homogenates Western blotted for GGA and AP-1. In both fly lines depletion of GGA was achieved to >95%. Note that AP-1 expression does not alter in the GGA knockdown flies. (B) GGA knockdown flies were dissected into head and testes and the efficiency of knockdown determined by Western blotting. Note that the GGA antibody recognises a doublet, but only the bottom band is knocked down (arrowhead). Quantitation of the gels bands demonstrates that the knockdown efficiency in testes is >95%, similar to that seen in head extracts. Testes squashes from control (C) and GGA knockdown flies (D) were labelled with antibodies against GGA. Note that GGA labelling is almost completely lost in knockdown fly testes suggesting efficient GGA depletion. Bar, 20 μm.
Hirst and Carmichael BMC Cell Biology 2011 12:22 doi:10.1186/1471-2121-12-22