Drosophila Wing Disc Immunostaining

Primary antibodies used were mouse-anti-Wingless (4D4-s; 1:120; DSHB, University of Iowa); mouse-anti-Patched (Apa1-s; 1:40; DSHB, University of Iowa); rat-anti-DE-cadherin (DCAD2 concentrate; 1:200; DSHB, University of Iowa); rabbit-anti-GFP (1:1000, Abcam ab6556, Lot:GR3404234-1); rabbit-anti-Phospho-Histone H3 (PHH3, 1:1000, Cell Signaling #9701); rabbit-anti-Mmp2 (1:500, from ref. ^{67 (link)}); rabbit-anti-Vkg (1/500, from ref. ^{68 (link)}).
Tissue outlines were marked by Alexa Fluor 660 Phalloidin (1:100, A 22285, Sigma-Aldrich) which was added together with the other secondary antibodies. Secondary antibodies used were Alexa 488 (donkey anti-mouse A21202, donkey anti-rabbit A21206), Alexa 568 (donkey anti-mouse A10037, donkey anti-rabbit A10042, goat anti-rat A11077) and Alexa 647 (donkey anti-rabbit A31573) from Invitrogen and Alexa 647 (donkey anti-mouse 715 605 151) from Jackson ImmunoResearch. Secondary antibodies were used at 1:500 dilution. Discs were blocked in 2% normal donkey serum (017-000-121, Jackson ImmunoResearch).

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Harmansa S., Erlich A., Eloy C., Zurlo G, & Lecuit T. (2023). Growth anisotropy of the extracellular matrix shapes a developing organ. Nature Communications, 14, 1220.

Publication 2023

ab6556 Alexa 488 Alexa 568 Alexa 647

Alexa 568 Antibodies Cadherin Dilution Donkey Goat Histone h3 Lot 1 Mmp2 Mouse Phalloidin Rabbit Serum Tissue

Corresponding Organization : Collège de France

Other organizations : Centre National de la Recherche Scientifique, Aix-Marseille Université, Centrale Marseille, Institut de Recherche sur les Phénomènes Hors Équilibre, Ollscoil na Gaillimhe – University of Galway

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Variable analysis

independent variables

Primary antibodies used were mouse-anti-Wingless (4D4-s; 1:120; DSHB, University of Iowa)
Mouse-anti-Patched (Apa1-s; 1:40; DSHB, University of Iowa)
Rat-anti-DE-cadherin (DCAD2 concentrate; 1:200; DSHB, University of Iowa)
Rabbit-anti-GFP (1:1000, Abcam ab6556, Lot:GR3404234-1)
Rabbit-anti-Phospho-Histone H3 (PHH3, 1:1000, Cell Signaling #9701)
Rabbit-anti-Mmp2 (1:500, from ref. 67 (link))
Rabbit-anti-Vkg (1/500, from ref. 68 (link))

dependent variables

Not explicitly mentioned

control variables

Alexa Fluor 660 Phalloidin (1:100, A 22285, Sigma-Aldrich) which was added together with the other secondary antibodies
Secondary antibodies used were Alexa 488 (donkey anti-mouse A21202, donkey anti-rabbit A21206), Alexa 568 (donkey anti-mouse A10037, donkey anti-rabbit A10042, goat anti-rat A11077) and Alexa 647 (donkey anti-rabbit A31573) from Invitrogen and Alexa 647 (donkey anti-mouse 715 605 151) from Jackson ImmunoResearch. Secondary antibodies were used at 1:500 dilution
Discs were blocked in 2% normal donkey serum (017-000-121, Jackson ImmunoResearch)

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