Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc

Abstract

Summary: Apicobasal polarity determinants are potential tumor suppressors that have been extensively studied. However, the precise mechanisms by which their misregulation disrupts tissue homeostasis are not fully understood. Here, we present a comprehensive protocol for establishing a conditional RNAi knockdown of scribble in Drosophila wing imaginal disc. We describe steps for generating fly lines, conditional knockdown in host stocks, and sample preparation. We then detail procedures for imaging, image analysis, and verification of wing disc phenotypes by various antibodies.For complete details on the use and execution of this protocol, please refer to Huang et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Developmental Biology
• Genetics
• Microscopy
• Model Organisms
• Molecular Biology
• Gene Expression