Journal: bioRxiv

Article Title: A genome engineered tool set for Drosophila TGF-β/BMP signaling studies

doi: 10.1101/2024.07.02.601693

Figure Lengend Snippet: (A) Overview of the HA tools comprising FB-GFP, deGradHA, GrabHA Ext , GrabHA Int and GrabHA-ECM. NSlmb = N-terminal part of Slmb, CDS = coding sequence, SP = signal peptide, TM = transmembrane domain, aa = amino acids. (B) anti-pMad and anti-Brk staining of wing discs expressing HA-Dpp, GrabHA Ext in dpp producing cells or a combination of both. Anterior-posterior compartment boundary is marked by red dashed line. Scale bars: 50 μm. (C) Anti-Brk and anti-BrdU staining of wing discs expressing HA-or YFP-tagged versions of Brk either alone or in combination with the indicated tool in the dorsal compartment using ap-Gal4. Red dashed lines mark dorso-ventral compartment boundary. Adult wings carrying HA-or YFP-tagged versions of Brk either alone or in combination with overexpression of GrabHA Int or iGFPi throughout the wing pouch using nub-Gal4. Scale bars: 50 μm (wing discs), 250 μm (adult wings).

Article Snippet: The following antibodies were used in this study: chicken anti-GFP (1:1000, Abcam), rat anti-HA (1:200, Roche), mouse anti-Dlg (1:50, DSHB), rabbit anti-pMad (1:500, Abcam), rabbit anti-pMad (1:500, Cell Signalling), guinea pig anti-Brk (1:500, Hilary Ashe), mouse anti-Wg (1:40, DSHB), mouse anti-Ptc (1:40, DSHB), rat anti-DCad (1:50, DSHB), mouse anti-Hts (1:10, DSHB), mouse anti-Brdu antibody (1:100, BD Bioscience), Alexa fluorophore-conjugated secondary antibodies (1:500; Thermo Fisher Scientific) and Hoechst 33342 (1:5000; Invitrogen).

Techniques: Sequencing, Staining, Expressing, BrdU Staining, Over Expression