Computational protocol: Dysfunction of Wntless triggers the retrograde Golgi-to-ER transport of Wingless and induces ER stress

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Protocol publication

[…] Antibody staining of wing imaginal discs or cells was performed using standard protocols. The following primary antibodies were used: rat anti-Bip (MAC143, Abcam), mouse anti-Wg (4D4; DSHB), guinea pig anti-Sens, rabbit anti-Wls, rabbit anti-GFP Alexa Fluor 488 (Molecular Probe), mouse anti-lacZ (Abmart), Rat anti-KDEL (Abcam), and Rabbit anti-GM130 (Abcam). The nuclei were stained by Hoechst 33258 (Sigma). The primary antibodies were detected by fluorescent-conjugated secondary antibodies from Jackson ImmunoResearch Laboratories. Confocal images were collected using a Lecia TCS SP5 confocal microscope with 40X/1.25 oil objectives. Adult thoracic bristle and wing images were obtained using a Nikon SMZ1500 microscope. Z-section images were taken with a Zeiss LSM 780 confocal microscope with 40X/1.3 oil objectives for the colocalization analysis. For determination of the Pearson’s correlation coefficient, the JACoP plugin for ImageJ was applied. Images were processed with ImageJ and Adobe Photoshop. […]

Pipeline specifications

Software tools Coloc, JACoP, ImageJ
Applications Laser scanning microscopy, Microscopic phenotype analysis
Organisms Drosophila melanogaster, Homo sapiens