Skip to main content
. 2004 Aug 1;18(15):1812–1823. doi: 10.1101/gad.298004

Figure 4.

Figure 4.

DNOS4 is endogenously expressed in Drosophila and forms complexes with DNOS1. (A) DNOS4 is endogenously expressed in Drosophila. Head protein extracts from wild-type and transgenic GMR-DNOS4-FLAG flies were analyzed by immunoblotting using DNOS4-specific antibody (anti-DNOS4 Ab) or the preimmune serum. Control lysates from nontransfected 293 cells (293) and from cells transfected with the pDNOS4 expression construct without FLAG tag (293-DNOS4) were probed using the same antibody or the preimmune serum. In addition to the ectopically expressed DNOS4-FLAG protein (86 kDa), which was detected in head extracts of the GMR-DNOS4-FLAG transgenic flies only, DNOS4-specific antibody immunoreacted with 84 kDa protein, which was expressed both in wild-type and in transgenic flies, and had the same mobility as the DNOS4 protein transiently expressed in 293 cells (lane 293-DNOS4). (B) In vivo complex formation between the endogenous DNOS1 and DNOS4 proteins. Protein complexes were immunoprecipitated from wild-type embryo extracts using either DNOS1- or DNOS4-specific antibody (Ab for immunoprecipitation) and cross-analyzed by immunoblotting with either DNOS1- or DNOS4-specific antibody (Ab for immunoblotting). The antibodies used for immunoprecipitation and for immunoblotting are indicated. (C–J) Expression of DNOS1 (green signal) and DNOS4 (red signal) in embryos (C–F), third-instar leg disc (G), pupae (∼3 h after puparium formation) leg disc (H), third-instar wing disc (I), and third-instar eye disc (J). (F) Embryos carrying a homozygous deletion of the dNOS locus were used as a control for the specificity of the antibodies.