Figure 6. Ubiquitous but not tissue specific knockdown of itpr recapitulates itpr^sv35/ug3 phenotypes.

(A) Act5c dsitpr larvae (Act5cGAL4/UASdsitpr¹⁰⁶³) did not undergo pupation and died as 3^rd instars. The number of pupae formed in Dilp2 dsitpr (Dilp2GAL4/UASdsitpr¹⁰⁶³; UASdicer/+), Ddc dsitpr (DdcGAL4/UASdsitpr¹⁰⁶³; UASdicer/+) and C155 dsitpr (Elav^C155GAL4; UASdsitpr¹⁰⁶³/+;UASdicer/+) was similar to controls (not significant; p>0.05; Student's t-test). GFP positive larvae were used as controls in each RNAi experiment. Three batches of 25 2^nd instar larvae were screened for each of the indicated genotypes. Results are expressed as mean±SEM. (B) Significant reduction in larval size was observed in Actin5cGAL4/UASdsitpr¹⁰⁶³ 3^rd instars (∼120 hrs AEL) compared to controls (Actin5cGAL4 or UASdsitpr¹⁰⁶³/CyoG). (C) RT-PCR gel with up-regulation of dLipase-3 transcript levels in RNA isolated from Actin5cGAL4/UASdsitpr¹⁰⁶³ larvae (∼ 85 hrs AEL) compared to RNA from controls. (D) A western blot with reduced InsP₃R (280Kda) protein levels in lysates from Actin5cGAL4/UASdsitpr¹⁰⁶³ 3^rd instar (∼120 hrs AEL) larvae as compared to lysates from controls. Equal levels of the loading control α-spectrin (278Kda) confirm that similar quantities of protein lysates were loaded in each lane.