Figure 8. dLKR/SDH is dynamically expressed during development, genetically interacts with EcR/Usp to control the timing of transcription, and influences wing development. (A) Expression of dLKR/SDH during development. Real-time PCR was performed in duplicate, and the mRNA expression levels were normalized against the internal control gene rp49. The error bars represent SEM. WPP, white prepupae. (B) Expression of dLKR/SDH during salivary gland programmed cell death. Real-time PCR was performed as in A using salivary gland cDNA at the indicated stages of development. −2, 0, and +2 refer to hours after head eversion (hRHE), corresponding to the endogenous ecdysone pulse. (C) 20 μg of salivary gland lysates from animals at 0 h after head eversion were analyzed by immunoblotting using the dLKR/SDH antibody. Salivary glands from wild type (WT) and those expressing salivary gland (sgs3-GAL4)–specific dLKR/SDH transgenic (dLKR/SDH) or dLKR/SDH double-stranded RNA (RNAi) were used. dLKR/SDH bands are indicated by an arrow. The asterisk indicates a nonspecific protein of ∼65 kD detected by the dLKR/SDH antibody. (D) Real-time PCR was performed as in A using salivary gland cDNA from animals as in C. (E) Effects of dLKR/SDH on dronc expression in salivary glands. Real-time PCR was performed as in A. The developmental stages are indicated as hours after head eversion. (F) Wing size was decreased in homozygous (Df/Df) adult females compared with wild-type controls. Knockdown of dLKR/SDH in the posterior wing compartment using en-GAL4 (en>RNAi2) resulted in a smaller wing compared with the control, en-GAL4 (en). (G) Wing areas as measured using the histogram tool in Photoshop. Df/Df adult females had a significantly smaller wing area than wild-type and Df/+ control animals (P < 0.0001). dLKR/SDH RNAi lines 1 and 2 expressed using en-GAL4 also resulted in a significantly smaller wing (P = 0.0006 and P < 0.0001 for RNAi line 1 and line 2, respectively). Interaction with the EcRF645A dominant-negative allele alleviated this decrease in size. Significance was calculated using a t test. (H) Genetic interaction tests using GMR-GAL4–driven expression of dLKR/SDH RNAi and EcRF645A (GMR>EcR;RNAi) in the eye resulted in suppression of the characteristic scar (arrow) caused by EcRF645A expression (GMR>EcR).
Figure 8.

dLKR/SDH is dynamically expressed during development, genetically interacts with EcR/Usp to control the timing of transcription, and influences wing development. (A) Expression of dLKR/SDH during development. Real-time PCR was performed in duplicate, and the mRNA expression levels were normalized against the internal control gene rp49. The error bars represent SEM. WPP, white prepupae. (B) Expression of dLKR/SDH during salivary gland programmed cell death. Real-time PCR was performed as in A using salivary gland cDNA at the indicated stages of development. −2, 0, and +2 refer to hours after head eversion (hRHE), corresponding to the endogenous ecdysone pulse. (C) 20 μg of salivary gland lysates from animals at 0 h after head eversion were analyzed by immunoblotting using the dLKR/SDH antibody. Salivary glands from wild type (WT) and those expressing salivary gland (sgs3-GAL4)–specific dLKR/SDH transgenic (dLKR/SDH) or dLKR/SDH double-stranded RNA (RNAi) were used. dLKR/SDH bands are indicated by an arrow. The asterisk indicates a nonspecific protein of ∼65 kD detected by the dLKR/SDH antibody. (D) Real-time PCR was performed as in A using salivary gland cDNA from animals as in C. (E) Effects of dLKR/SDH on dronc expression in salivary glands. Real-time PCR was performed as in A. The developmental stages are indicated as hours after head eversion. (F) Wing size was decreased in homozygous (Df/Df) adult females compared with wild-type controls. Knockdown of dLKR/SDH in the posterior wing compartment using en-GAL4 (en>RNAi2) resulted in a smaller wing compared with the control, en-GAL4 (en). (G) Wing areas as measured using the histogram tool in Photoshop. Df/Df adult females had a significantly smaller wing area than wild-type and Df/+ control animals (P < 0.0001). dLKR/SDH RNAi lines 1 and 2 expressed using en-GAL4 also resulted in a significantly smaller wing (P = 0.0006 and P < 0.0001 for RNAi line 1 and line 2, respectively). Interaction with the EcRF645A dominant-negative allele alleviated this decrease in size. Significance was calculated using a t test. (H) Genetic interaction tests using GMR-GAL4–driven expression of dLKR/SDH RNAi and EcRF645A (GMR>EcR;RNAi) in the eye resulted in suppression of the characteristic scar (arrow) caused by EcRF645A expression (GMR>EcR).

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal