Cellular Localization of Lv-AE3 mRNA in Gill by ISH

The cellular localization of Lv-AE3 mRNA was performed in the gill with high transcript levels by ISH as described previously (Ruan et al., 2020 (link)). The digoxigenin (DIG)-dUTP-labeled DNA probe targeting Lv-AE3 was generated by using a DIG High Prime DNA Labeling and Detection Starter Kit II (Roche, Switzerland). The slices incubated by DIG-labeled probe were incubated with horseradish peroxidase (HRP)-conjugated anti-DIG antibody (Roche), and the ISH signal was developed by a diaminobenzidine (DAB, MXB Biotechnologies) reaction and the nucleus were stained with hematoxylin. The Lv-AE3 mRNA expressed cells in the gill were observed and recorded with a Leica DM-IRB light microscope (Leica, Germany).

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Zhang X., Yang H., Li H., Chen T., Ruan Y., Ren C., Luo P., Wang Y., Liu B., Li H., Zhong P., Zhang J., Jiang X, & Hu C. (2021). Molecular Identification of Anion Exchange Protein 3 in Pacific White Shrimp (Litopenaeus vannamei): mRNA Profiles for Tissues, Ontogeny, Molting, and Ovarian Development and Its Potential Role in Stress-Induced Gill Damage. Frontiers in Physiology, 12, 726600.

Publication 2021

DIG High Prime DNA Labeling and Detection Starter Kit II horseradish peroxidase (HRP)-conjugated anti-DIG antibody diaminobenzidine (DAB,

Anti antibody Digoxigenin Dna probe Dutp Gill Horseradish peroxidase Microscope light Mrna Nucleus

Corresponding Organization : Chinese Academy of Sciences

Other organizations : University of Chinese Academy of Sciences, Hebei University, Guangdong Laboratory Animals Monitoring Institute, Jinan University, Institute of Hydrobiology

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Variable analysis

independent variables

Probe targeting Lv-AE3 mRNA

dependent variables

Cellular localization of Lv-AE3 mRNA in the gill

control variables

Experimental procedure described in Ruan et al., 2020

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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