de Faria Isaque J. S., Imler Jean-Luc, Marques João T.
Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies Article de journal
Dans: STAR Protocols, vol. 4, iss. 1, 2023.
Résumé | Liens | BibTeX | Étiquettes: antibody, cell bioloby, imler, Immunology, M3i, Marques, microbiology, Microscopy, Molecular Biology
@article{deFaria2023,
title = {Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies},
author = {Isaque J.S. de Faria and Jean-Luc Imler and João T. Marques},
url = {https://doi.org/10.1016/j.xpro.2022.102033},
doi = {10.1016/j.xpro.2022.102033},
year = {2023},
date = {2023-03-17},
urldate = {2023-03-17},
journal = {STAR Protocols},
volume = {4},
issue = {1},
abstract = {Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissues and other organisms. We describe the procedures to prepare insect cells for viral infection, followed by RNA extraction and in vitro production of synthetic dsRNA controls. We then detail the steps for dsRNA detection by immunoblotting and immunofluorescence. For complete details on the use and execution of this protocol, please refer to de Faria et al. (2022).1},
keywords = {antibody, cell bioloby, imler, Immunology, M3i, Marques, microbiology, Microscopy, Molecular Biology},
pubstate = {published},
tppubtype = {article}
}
Prakash Pragya, Roychowdhury-Sinha Arghyashree, Goto Akira
Verloren negatively regulates the expression of IMD pathway dependent antimicrobial peptides in Drosophila Article de journal
Dans: Scientific Reports, vol. 11, no. 15549, 2021.
Résumé | Liens | BibTeX | Étiquettes: bacteria, Biochemistry, DNA, Fungi, Gene Expression, gene regulation, Genetics, hoffmann, Immunochemistry, Immunology, infection, inflammation, Innate immune cells, innate immunity, M3i, microbiology, Molecular Biology, pathogens, RNA, RNAi, Signal Transduction, Transcription
@article{Goto2021,
title = {Verloren negatively regulates the expression of IMD pathway dependent antimicrobial peptides in Drosophila},
author = {Pragya Prakash and Arghyashree Roychowdhury-Sinha and Akira Goto},
url = {https://www.nature.com/articles/s41598-021-94973-0},
doi = {10.1038/s41598-021-94973-0},
year = {2021},
date = {2021-07-30},
journal = {Scientific Reports},
volume = {11},
number = {15549},
abstract = {Drosophila immune deficiency (IMD) pathway is similar to the human tumor necrosis factor receptor (TNFR) signaling pathway and is preferentially activated by Gram-negative bacterial infection. Recent studies highlighted the importance of IMD pathway regulation as it is tightly controlled by numbers of negative regulators at multiple levels. Here, we report a new negative regulator of the IMD pathway, Verloren (Velo). Silencing of Velo led to constitutive expression of the IMD pathway dependent antimicrobial peptides (AMPs), and Escherichia coli stimulation further enhanced the AMP expression. Epistatic analysis indicated that Velo knock-down mediated AMP upregulation is dependent on the canonical members of the IMD pathway. The immune fluorescent study using overexpression constructs revealed that Velo resides both in the nucleus and cytoplasm, but the majority (~ 75%) is localized in the nucleus. We also observed from in vivo analysis that Velo knock-down flies exhibit significant upregulation of the AMP expression and reduced bacterial load. Survival experiments showed that Velo knock-down flies have a short lifespan and are susceptible to the infection of pathogenic Gram-negative bacteria, P. aeruginosa. Taken together, these data suggest that Velo is an additional new negative regulator of the IMD pathway, possibly acting in both the nucleus and cytoplasm.},
keywords = {bacteria, Biochemistry, DNA, Fungi, Gene Expression, gene regulation, Genetics, hoffmann, Immunochemistry, Immunology, infection, inflammation, Innate immune cells, innate immunity, M3i, microbiology, Molecular Biology, pathogens, RNA, RNAi, Signal Transduction, Transcription},
pubstate = {published},
tppubtype = {article}
}