Figure 6

Glutamate receptor subunit mRNA and protein is not reduced in sec8 mutants. A: Relative mRNA levels for glutamate receptor subunits GluRIIA (left), GluRIIB (middle), and GluRIIC (right), as measured by quantitative real-time RT-PCR in control and sec8 mutant first instar larvae. B: Example immunoblot from first instar larvae (24–28 h AEL) probed with a anti-GluRIIB antibody. The arrow points to the GluRIIB band. GluRIIA immunoblots have been previously described [71]. GluRIIA and GluRIIB bands are eliminated in Df(2L)SP22 mutants, in which GluRIIA and GluRIIB genes are deleted (not shown). Loading control was total protein; to ensure that the same amount of protein was loaded for both control and mutant, total extracted protein quantity was measured using a Bradford assay and then the same amount of protein was loaded on to the gel for each mutant (either 20 μg for GluRIIB blots or 50 μg for GluRIIA blots). Loading was also checked with Coomassie staining (not shown). C: Quantification of total GluRIIA (left) and GluRIIB (right) protein, as measured from immunoblots such as the one shown in B.