Lung mononuclear cells miRNA analysis

miRNA analysis was carried out as described previously (Alharris et al., 2018 (link); Neamah et al., 2019 (link)). Briefly, total RNA, including miRNA, was isolated from lung mononuclear cells using the miRNeasy kit from QIAGEN and following the protocol of the company. Microarray was performed using Affymetrix miRNA Array (version 4.1). Raw files generated from the miRNA microarray were uploaded to Gene Expression Omnibus (http://www.ncbi.nlm.nih.gov/geo) and deposited under accession number GSE220159. By using Transcriptome Analysis Console (TAC, ThermoFisher, United States), Log2 fold change of more than 3,000 miRNAs was detected from the raw array data, and only those miRNAs that were altered more than 2-fold were considered for further analysis. Filtered miRNAs were analyzed for their role in various biological pathways using Ingenuity Pathway Analysis (IPA) software http://www.ingenuity.com (Qiagen, Germany). Also, microRNA.org database was used to examine the sequence alignment regions between miR-100-5p and its targeted genes.

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Alghetaa H., Mohammed A., Singh N., Wilson K., Cai G., Putluri N., Nagarkatti M, & Nagarkatti P. (2023). Resveratrol attenuates staphylococcal enterotoxin B-activated immune cell metabolism via upregulation of miR-100 and suppression of mTOR signaling pathway. Frontiers in Pharmacology, 14, 1106733.

Publication 2023

miRNeasy kit Transcriptome Analysis Console (TAC,

Biological Cells Gene expression Genes Lung Microarray Microrna Sequence alignment Transcriptome analysis

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Variable analysis

independent variables

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dependent variables

Expression levels of miRNAs

control variables

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controls

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Negative control: Not mentioned

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