(A) Illustration of the late stage larvae (L3), body is organized into repeated left-right, mirror image hemisegments. Muscles (in gray) have a stereotyped pattern. (B) Illustration of muscles in a …
Test for difference in the number of synaptic branches on ventral muscles in NB3-1>Hb compared to Control.
(A) Illustration of NB3-1 lineage progression. Each gray arrowhead represents cell division. Large circles are neuroblasts and smaller circles are neurons. Abbreviations: MN is motor neuron, IN is …
(A–B) Illustrations of NB3-1 lineage progression. Each gray arrowhead represents cell division. Each gray arrowhead represents cell division. Large circles are neuroblasts, and smaller circles are …
Test for difference in the number of embryonic RP MN molecular identities in NB3-1>Hbcompared to Control.
(A) Illustration of subcellular localization of neuromuscular synapse markers. Brunchpilot labels active zones, Discs large is a scaffolding protein strongly localized at post-synapse, and GluRIIA …
(A) Illustration of neuromuscular junction (NMJ) electrophysiology approach on third instar larvae (L3). Abbreviation NT is neurotransmitter. (B) Illustration highlighting (darkened) Muscle 30 whose …
Test for the difference in EPSP, mEPSP, and Quantal Content for muscle 14 and 6 in NB3-1>Hb compared to Control.
(A–B) Illustrations of NB7-1 lineage progression. Each gray arrowhead represents cell division. Each gray arrowhead represents cell division. Large circles are neuroblasts, and smaller circles are …
Test for difference in the number of synaptic branches on dorsal muscles in NB7-1>Cas compared to Control.
(A–B) Illustrations of NB7-1 lineage progression. Each gray arrowhead represents cell division. Each gray arrowhead represents cell division. Large circles are neuroblasts, and smaller circles are …
(A–C) Illustration of NB7-1 lineage progression. Each gray arrowhead represents cell division. Each gray arrowhead represents cell division. Large circles are neuroblasts, and smaller circles are …
Test for difference in the number of synaptic branches on dorsal muscles in NB7-1>Pdm compared to Control.
(A) Illustration of NB7-1 lineage progression. Each gray arrowhead represents cell division. Each gray arrowhead represents cell division. Large circles are neuroblasts, and smaller circles are …
(A) Illustration of subcellular localization of neuromuscular synapse markers. Brunchpilot labels active zones, Discs large is a scaffolding protein strongly localized at post-synapse, GluRIIA is a …
(A–D) Illustration of calcium imaging protocol and analysis are shown. See Materials and methods for details. (E–H) Images of calcium signals on L3 Muscle 4. Control is NB7-1-GAL4/+; …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (D. melanogaster) | DIP-alpha-GAL4 | Robert Carrillo (UChicago) | ||
Genetic reagent (D. melanogaster) | ac:VP16, gsb:v8v (aka NB7-1-GAL4) | Minoree Kohwi (Columbia) | ||
Genetic reagent (D. melanogaster) | MHC-CD8-GCaMP6f-Sh | Bloomington stock center [BL] 67739 | BDSC Cat# 67739, RRID:BDSC_67739 | |
Genetic reagent (D. melanogaster) | UAS-Hb; UAS-Hb/TM2 | Bloomington stock center [BL] 32198 | BDSC Cat# 32198, RRID:BDSC_32198 | |
Genetic reagent (D. melanogaster) | 5172 J-GAL4 (aka NB3-1-GAL4) | Haluk Lacin (Washington University) | ||
Genetic reagent (D. melanogaster) | w1118 | Bloomington stock center [BL] 36005 | BDSC Cat# 36005, RRID:BDSC_36005 | |
Genetic reagent (D. melanogaster) | UAS-Cas | Chris Doe (University of Oregon) | ||
Genetic reagent (D. melanogaster) | UAS-HA-pdm2/CyO; UAS-HA-pdm2/TM3 | Chris Doe (University of Oregon) | ||
Genetic reagent (D. melanogaster) | UAS-myr-GFP | Bloomington stock center [BL] 32198 | BDSC Cat# 32198, RRID:BDSC_32198 | |
Antibody | rabbit anti-Eve (polyclonal) | Heckscher Lab | 1:1000 | |
Antibody | rabbit anti-Castor (polyclonal) | Chris Doe (Oregon) | 1:1000 | |
Antibody | guinea pig anti-Hb (polyclonal) | John Rientz (UChicago) | 1:1000 | |
Antibody | guinea pig anti-Kruppel (polyclonal) | John Rientz (UChicago) | 1:1000 | |
Antibody | Mouse anti-Eve (monoclonal) | 3C10 | DSHB Cat# 3C10 Anti-even skipped RRID:AB_528229 | 1:50 |
Antibody | chicken anti-GFP (polyclonal) | Aves #GFP-1020 | Aves Labs Cat# GFP-1020, RRID:AB_10000240 | 1:1000 |
Antibody | rat anti-Worniu (polyclonal) | Abcam #ab196362 | 1:250 | |
Antibody | mouse anti-En (monoclonal) | 4D9 | DSHB Cat# 4D9 anti-engrailed/invected, RRID:AB_528224 | 1:5 |
Antibody | guinea pig anti-Runt (polyclonal) | John Rientz (UChicago) | 1:500 | |
Antibody | guinea pig anti-HB9 (polyclonal) | Heather Broiher (Case Western) | 1:1000 | |
Antibody | mouse anti-Cut (monoclonal) | 2B10 | DSHB Cat# 2B10 anti-Cut homeobox RRID:AB_528186 | 1:50 |
Antibody | rat anti-Zfh2 (polyclonal) | Chris Doe (Oregon) | 1:800 | |
Antibody | rabbit anti-Smad3 (pMad) (polyclonal) | Abcam #52903 | Abcam Cat# ab52903, RRID:AB_882596 | 1:300 |
Antibody | mouse anti-Brp (monoclonal) | NC82 | Creative Diagnostics Cat# DMAB9116MD, RRID:AB_2392664 | 1:50 |
Antibody | mouse anti-DLG (monoclonal) | 4F3 | DSHB Cat# 4F3 anti-discs large, RRID:AB_528203 | 1:500 |
Antibody | mouse anti-GluRIIA (monoclonal) | 8B4D2 | DSHB Cat# 8B4D2 (MH2B), RRID:AB_52826 | 1:25 |
Antibody | Cy3-HRP (polyclonal) | Jackson ImmunoResearch 123-025-021 | Jackson ImmunoResearch Labs Cat# 123-025-021, RRID:AB_2338954 | 1:400 |
Antibody | 647-Phallion (polyclonal) | Thermofisher A22287 | Thermo Fisher Scientific Cat# A22287, RRID:AB_2620155 | 1:600 |