Figure 7

Depletion of Brm by RNAi in vivo affects the relative abundances of alternative transcripts from the Gpdh and lola genes in larvae. hs-GAL4 virgin females were crossed with UAS-BrmRNAi males, and early third instar larvae from the cross were heat shocked for 2 h at 37°C. Control experiments were carried out in parallel by crossing hs-GAL4 virgin females with wild type males W1118. (A) The effect of the depletion was analyzed 24 h after the heat shock by Western blotting using an anti-Brm antibody. The same blot was probed in parallel with an anti-tubulin antibody that served as a loading control. (B-E) hs-GAL4 virgin females were crossed with UAS-BrmRNAi males, the third instar larvae from the cross were heat shocked as above, and total RNA was isolated 24 h after the heat shock. Total RNA was purified and the abundances of selected transcripts were analyzed by RT-qPCR using the same primers as in Figures 3-6. In (B), (C) and (E), the relative abundance of each alternative transcript was calculated relative to the abundance of a constitutive exon. In (D), the relative abundance of each alternative transcript was calculated relative to the abundance of the Act5C mRNA, as in Figure 5. The results are expressed as average factor change between BrmRNAi and W1118 flies. The bars represent averages and the error bars represent standard deviations from two independent experiments, each quantified in duplicate (n = 4).