Journal: bioRxiv

Article Title: Executioner caspase is proximal to Fasciclin 3 which facilitates non-lethal activation in Drosophila olfactory receptor neurons

doi: 10.1101/2023.07.20.549821

Figure Lengend Snippet: (A) Representative expression patterns of Drice (Streptavidin (SA); magenta) and Fasciclin 3 (Fas3, anti-Fasciclin 3 antibody staining; green) in the adult male brains. Scale bar: 50 µm. (B) Representative expression patterns of Drice (SA; magenta) and Fas3 (anti-GFP antibody staining; green) in the antennal lobes (ALs) of the adult male brains. Scale bar: 50 µm. (C) Gene structure of Fas3. CDS (orange) and UTR (grey) are shown in boxes. (D) Schematic structures of Fas3 protein isoforms. Intracellular regions differ from each other. A peptide region used to raise anti-Fas3G antibody is shown. (E) IDR predictions of intracellular regions of each protein isoform. (F) Western blot of proteins extracted from adult heads. Anti-Fas3G antibody specifically detects Fas3 isoform G. (G) Western blot of adult male heads. Drice proximal proteins were purified using NeturAvidin. (H) Western blot of adult head proteins. Drice proximal proteins were purified using NeturAvidin.

Article Snippet: The primary antibodies used were mouse anti-Fasciclin 3 (7G10) antibody (1:20, #AB_528238, DSHB), rat anti-GFP (GF090R) monoclonal antibody (1:100, #04404-26, nacalai Tesque), mouse anti-FLAG M2 monoclonal antibody (1:500, F1804, Sigma), and rat anti-mCD8 monoclonal antibody (1:50, #MCD0800, Invitrogen).

Techniques: Expressing, Staining, Western Blot, Purification

Journal: bioRxiv

Article Title: DamID transcriptional profiling identifies the Snail/Scratch transcription factor Kahuli as Alk target in the Drosophila visceral mesoderm

doi: 10.1101/2021.01.25.428051

Figure Lengend Snippet: A. Kah belongs to the Snail/Scratch family of transcription factors sharing 5 zinc-finger domains. Schematic indicates the overall domain structure of the Drosophila family members, Kahuli, Snail, Escargot, Wornoi, CG12605 and Scratch. A’. Phylogenetic tree indicating the relationship of between Kah and the members of the Snail/Scratch family. B. Violin plots from scRNA-seq of wild-type embryos indicates Kah transcript is expressed in the embryonic VM and SM. C. Violin plots from scRNA-seq analysis of FACS sorted Hand-GFP expressing cells reveals expression of Kah mRNA in visceral, but not cardiac, mesoderm. D. Kah transcripts are abundant in SM and VM during embryogenesis, with increased expression levels in the visceral founder cell (FC) row. FCM, fusion competent myoblasts; sm, somatic musculature; vm, visceral musculature. E. Ectopic expression of jeb results in an increase of Kah expression in visceral FCMs. Conversely, animals devoid of Jeb/Alk signalling ( jeb weli mutants) lack the strong FC-specific Kah expression in the VM while SM expression remains unaltered. F-G’. A Kah.GFP gene duplication construct can be detected from stage 10 embryos in the VM, with no clear distinction between FCs (marked by rp298-lacZ, red, inset depicts a close-up in LUT colors) and FCMs. Lateral view (F, F’), dorsal view (G, G’). H-H’. After myoblast fusion (stage 13), Kah.GFP is still maintained in visceral (vm) and somatic musculature (sm). Dorsal view. I-J’. Expression of endogenously-tagged Kah Cterm.OLLAS is similar to Kah.GFP, but appears to be enriched in visceral FCs (marked by Org-1, green, inset depicts a close-up in LUT colors). Lateral view (I, I’), dorsal view (J, J’). K-K’. Stage 13 embryos show Kah Cterm.OLLAS both in the visceral and somatic muscles (vm and sm, respectively). Dorsal view.

Article Snippet: Primary antibodies used were: guinea pig anti-Alk (1:1000 ( )), rabbit anti-Alk (1:750 ( )), chicken anti-β-galactosidase (1:200; Abcam ab9361), mouse anti-Fasciclin3 (1:50; DSHB 7G10), mouse anti-Antp (1:50; DSHB 4C3) rabbit anti-GFP (1:500; Abcam ab290), chicken anti-GFP (1:300; Abcam ab13970), mouse anti-Wg (1:50, DSHB 4D4), rat anti-OLLAS (1:200, pre-absorbed on w 1118 embryos; Abnova), rabbit anti-Org-1 (1:1000, ( )), sheep anti-digoxygenin-AP fab fragment 1:4000 (Roche).

Techniques: Expressing, Construct, Muscles

Journal: bioRxiv

Article Title: DamID transcriptional profiling identifies the Snail/Scratch transcription factor Kahuli as Alk target in the Drosophila visceral mesoderm

doi: 10.1101/2021.01.25.428051

Figure Lengend Snippet: A. Schematic overview of the newly generated Kah alleles: Kah Cterm.OLLAS , Kah ΔATG and Kah ΔZnf , together with the Kah f06749 PiggyBac insertion allele. B. Stage 10 embryos exhibit FC-specific expression of Org-1 (red) and HandC-GFP (green) FC markers. Dorsal views. C. Wild-type embryos at stage 16 are characterized by three midgut constrictions while Kah mutants fail to form the first midgut constriction. Kah mutants express Wg and dpp at levels comparable with control ( w 1118 ) embryos. Dorsal views. D. Quantification of the midgut constriction phenotype observed in C. E. pnt Δ88 mutants display a midgut constriction phenotype similar to that observed in Kah mutants. Dorsal views. F. Kah mutants display abnormal organisation of midgut musculature, visualized with the nuclear HandC-GFP reporter FC markers (green). Lateral views. G. Quantification of the number of nuclei present in wild-type ( w 1118 ) and Kah mutants.

Article Snippet: Primary antibodies used were: guinea pig anti-Alk (1:1000 ( )), rabbit anti-Alk (1:750 ( )), chicken anti-β-galactosidase (1:200; Abcam ab9361), mouse anti-Fasciclin3 (1:50; DSHB 7G10), mouse anti-Antp (1:50; DSHB 4C3) rabbit anti-GFP (1:500; Abcam ab290), chicken anti-GFP (1:300; Abcam ab13970), mouse anti-Wg (1:50, DSHB 4D4), rat anti-OLLAS (1:200, pre-absorbed on w 1118 embryos; Abnova), rabbit anti-Org-1 (1:1000, ( )), sheep anti-digoxygenin-AP fab fragment 1:4000 (Roche).

Techniques: Generated, Expressing, Control