Regulation of Drosophila Metamorphosis by Xenobiotic Response Regulators
Figure 5
Interrelationships between Ras signaling and CncC functions.
(A). Effects of CncC depletion on the time of pupation by larvae that expressed RasV12 in the PG. The proportion of larvae that had formed pupae was plotted as a function of the time after hatching for control larvae (phm>, black), larvae that expressed either RasV12 alone (phm>rasV12, red) or RasV12 together with the shRNA targeting CncC (phm>rasV12, cncC-RNAi, green) in the PG. The data represent the means and standard deviations from three repeats using 20–30 larvae in each. (B). Effects of CncC depletion on the sizes of pupae formed by larvae that expressed RasV12 in the PG. The lengths of pupae formed by the larvae described in part A were measured. The data represent the means and standard deviations of 30 pupae of each genotype (*, p<0.001). (C). Effects of CncC depletion and RasV12 expression in the PG on pupal development. The terminal stage of development was recorded for pupae formed by larvae that expressed either the shRNA targeting CncC alone (5015>cncC-RNAi), RasV12 alone (5015>rasV12), or RasV12 in combination with the shRNA targeting CncC (5015>rasV12, cncC-RNAi) in the PG. The proportion of pupae that arrested at early (P1–P9, open bar) and late (P10–P15, striped bar) pupal stages and that eclosed (adult, solid bar) are indicated (images shown on the right). Early and late stage pupae were distinguished by the absence or presence of red eye pigment (red arrow) and dark wings (blue arrow). Approximately 100 animals of each genotype were scored. (D). Effects of RasV12 expression in salivary glands on CncC occupancy on polytene chromosomes. Polytene chromosomes from control larvae (Sgs3>) and larvae that expressed RasV12 (Sgs3>rasV12) were stained using anti-CncC antibody. Selected loci whose occupancy by CncC changed upon RasV12 expression are labeled. (E). Effects of RasV12 and CncC fusion or CncC-RNAi expression in salivary glands on transcription of genes whose occupancy was affected by RasV12 expression and of control xenobiotic response genes. The levels of the transcripts indicated above the graphs were measured in salivary glands that expressed the proteins or the shRNA indicated. All transcript levels were normalized by the level of Rp49 transcripts. The data represent the means and standard deviations from two separate experiments (*, p<0.05).