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Dcp-1 is a laboratory product offered by Cell Signaling Technology. It is a protein that plays a role in cellular processes. The core function of Dcp-1 is to participate in the regulation of gene expression and cellular development, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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4 protocols using dcp 1

1

Immunohistochemical Staining of Drosophila Testes

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Immunohistochemical staining was performed as described previously with a few modifications (Lin et al., 2021 (link)). Drosophila adult testes were dissected in 1 × phosphate buffer saline (PBS) and fixed in 4% paraformaldehyde for 20 min. After washing with PBST (0.3% Triton in 1× PBS) three times, the testes were incubated in a blocking buffer (5% goat serum in 0.1% PBST) for 1 h. Next, the blocking buffer was replaced by primary antibodies mixed in PAT (1% BSA, 0.1% Triton and 0.01% sodium azide in 1× PBS) and the incubation lasted for overnight. Then, the testes were washed three times with 0.1% PBST and incubated in secondary antibodies mixed in 0.1% PBST for 1 h. After washing with 0.1% PBST three times, the tissues were stained with DAPI for 10 min and washed three times before mounting with Fluoromount-G. DAPI (#MBD0015, Sigma-Aldrich, Germany), DCP1 (#9578S, Cell signaling, USA).
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2

Immunohistochemistry Embryo Staining

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For immunohistochemistry embryos were fixed and stained according to standard procedures. Guinea pig anti-SIMU (30 (link)) and guinea pig anti-Drpr (32 (link)) were used at a 1:5000 and 1:100 concentrations, respectively. Rabbit anti-activated caspase 3 (Dcp-1) (Cell Signaling) and mouse anti-GFP (Roche) were used at 1:100 concentration. Rabbit anti-Crq antibody (1:500) is a gift from N. Franc. Rabbit anti-Srp antibody (1:100) is a gift from J. Casanova, K. Campbell and N. Martin. Rabbit anti-Peroxidasin antibody (1:2000) is a gift from Jiwon Shim. Fluorescent secondary antibodies (Cy3/and Cy5/Jackson ImmunoResearch; Alexa Fluor 488/Molecular Probes) were used at 1:200 dilutions. For TUNEL labeling embryos were re-fixed, washed and labeled with the In Situ Cell Death Detection kit (Roche) according to the manufacture instructions. Images were acquired on a confocal microscope Zeiss LSM 700 or on a Zeiss Axio Observer microscope equipped with an Apotome system using the AxioVision software. 75% Glycerol solution was used as the imaging medium.
Live imaging was carried out by dechorionating embryos (stage 15), mounting them under Halocarbon oil, injecting 2–3% egg volume of LysoTracker (Molecular Probes) as described in Ref. (33 (link)). Recording started 30 min following injection.
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3

Immunofluorescence Analysis of Eye Primordia

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Fixation and immunofluorescence of eye primordia was done as in [56 (link)]. Imaging was carried out on a Leica SPE confocal setup. Images were then processed using ImageJ. Primary antibodies used were rabbit anti-phospho-histone H3 (pH3) used at 1/1,000 (Sigma), rabbit against the cleaved (active) form of Dcp 1 (Dcp-1) used at 1/200 (ASP216, Cell Signaling Technology), and rabbit anti-cleaved (active) caspase 3 (Cas3*) used at 1/500 (D175, Cell Signaling Technology). Fluorescently labeled secondary antibodies were from Molecular Probes and used at 1/1,000. Rhodamine-phalloidin (Life Technologies) was used at 1/400 in some experiments to counterstain the tissue. DAPI (4′,6-diamidino-2-phenylindole, 1/10,000) was used in some experiments to counterstain nuclei. All primary and secondary antibodies were diluted in PBT (PBS + 0.1% Triton X-100).
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4

Immunofluorescence Staining of Drosophila Tissues

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EADs and wing imaginal discs dissected from third instar Drosophila larvae (7 days AEL) were fixed for 25 min with 4% paraformaldehyde in PBS containing 0.1% Triton X (PBS-T) at room temperature. Adult female intestines were fixed with 4% paraformaldehyde in PBS for 48 h at 4°C. Fixed tissues were washed three times with PBS-T, intestines in PBS only. Primary antibodies were diluted in blocking buffer (PBS-T with 0.3% BSA) and tissues were stained overnight at 4°C. The following primary antibodies were used: rabbit anti-Drosophila cleaved Death caspase 1 (Dcp-1, 1:500, Cell Signaling Technology Cat #9578, RRID:AB_2721060), rat-anti Ecd N-term (1:500, 22 (link)), mouse-anti Armadillo (Arm, 1:20, Developmental Studies Hybridoma Bank #N2 7A1; RRID:AB_528089), mouse-anti disc large (Dlg1, 1:100, Developmental Studies Hybridoma Bank #4F3; RRID:AB_528203). After washing, the samples were incubated with the corresponding Cy3- or Cy5-conjugated secondary antibodies (1:1000, Jackson ImmunoResearch Labs Cat# 711-175-152, RRID:AB_2340607, Cat# 712-165-150, RRID:AB_2340666, Cat# 715-165-151, RRID:AB_2315777) for 2 h at room temperature and counterstained with DAPI (1:1000 dilution of 5 mg/ml stock, Carl Roth GmbH Cat# 6335.1) to visualize nuclei. Tissues were mounted on glass slides in Dabco-Mowiol 4–88 (Sigma-Aldrich Cat# D2522 and Cat# 81381) and imaged within 72 h.
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