The purpose of this study was to preliminary investigate the function of the SOX18 gene in Hu sheep hair follicle dermal papilla cells.In this study,immunofluorescence staining was used to identify cells isolated from the hair follicles of Hu sheep.Subsequently,an overex-pression vector of the SOX 18 gene was conducted,and its overexpression efficacy was assessed using qRT-PCR and Western blot.The dermal papilla cells successfully identified and well-grown were divided into two groups.One group was transfected with SOX 18 overexpression vector,while the other was transfected with empty plasmid.Each group had 3 replicates.The total RNA was extracted from 6 samples using the Trizol method,followed by the construction of a cDNA library.The Illumina Novaseq6000 platform was used for sequencing.Subsequently,sample cor-relation and differentially expressed genes were analyzed.The differentially expressed genes were categorized based on GO and annotated using the KEGG pathway analysis to identify candidate genes and pathways associated with the regulation of the SOX 18 gene.To confirm the accuracy of the transcriptome data,9 differentially expressed genes were randomly chosen for qRT-PCR analysis.The immunofluorescence results revealed dermal papilla-related genes had high expres-sion in isolated cells,indicating a high purity of the isolated dermal papilla cells.qRT-PCR and Western blot results showed that overexpression of the SOX18 gene could increase the mRNA and protein levels of SOX18 in dermal papilla cells,indicating that the SOX18 overexpression vector could be used for subsequent experiments.Transcriptome analysis results showed a good correlation between samples.A total of 157 genes were differentially expressed between the SOX 18 overexpression group and the control group,of which 103 genes were up-regulated after SOX18 overexpression and 54 genes were down-regulated.qRT-PCR analysis showed high accu-racy of transcriptome data.GO functional analysis showed that some differential genes were enriched in cell progression and hair follicle development.KEGG enrichment analysis showed that some differentially expressed genes were enriched in signaling pathways related to cell prolif-eration and hair follicle development.GO functional analysis and KEGG enrichment analysis indicated that SOX18 played a role in the proliferation of dermal papilla cells and the development of hair follicles.In this study,transcriptome sequencing analysis found that SOX18 may affect the proliferation of dermal papilla cells and the development of hair follicles by affecting genes and signaling pathways related to cell proliferation and hair follicle development.The study results provide a theoretical basis for understanding the molecular regulatory mechanism of the SOX18 gene in Hu sheep dermal papilla cells and hair follicles.
维普
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