Transcriptional profiling of macrophages infected with Fonsecaea monophora

Abstract

Summary Fonsecaea monophora is a member of dematiaceous fungi capable of causing chromoblastomycosis through traumatic injury. However, little is known about the pathogenesis and early interactions between F. monophora and host. The aim of this study was to explore the potential mechanism of macrophages against F. monophora, especially the role of melanin during the pathogenic process. We carried out RNA sequencing based on the Illumina system. It showed that according to melanin contents, different strains of F. monophora induced different transcriptional profilings in macrophages. Functional analyses suggested the biological functions of differentially expressed genes were closely related to immune response, and the melanin might affect the interactions by regulating the MAPK signalling pathway of macrophages. Our results provide insights into the pathogenesis of infection by F. monophora conidia.

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“…RNA-seq was performed by Annoroad Gene Technology Co., Ltd. (Beijing, China). The RNA-seq methods were as described by Shi 56 . Total RNA was extracted from the testes collected at 0, 28, and 120 d from boars born in the CON group (designated N0, N28, N120) or LE group (designated L0, L28, and L120) using the TRIzol reagent (Invitrogen).…”

“…Total RNA was extracted from the testes collected at 0, 28, and 120 d from boars born in the CON group (designated N0, N28, N120) or LE group (designated L0, L28, and L120) using the TRIzol reagent (Invitrogen). Sequencing libraries were generated using the NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (#E7530L, NEB, USA) 56 .…”

“…The reference genomes were derived from the ENSEMBL database (http://www.ensembl.org/index.html), and clean data were mapped to the reference genome using HISAT2 (http://ccb.jhu.edu/software/hisat2/index.shtml) 56 . FPKM values were used to calculate the expression of genes in each sample.…”

“…FPKM values were used to calculate the expression of genes in each sample. Differential expression was analyzed by the DEGseq software (http://www.bioconductor.org/packages/release/bioc/html/DEGseq.html), Genes with q ≤ 0.05 and |log 2 ratio| ≥ 1 were considered as DEGs 56 .…”

“…GO terms with q < 0.05 were considered significantly enriched. For the functional classification of genes, by the same method as in the GO analysis, significantly enriched KEGG (http://www.kegg.jp/) pathways were identified 56 .…”