{"title":"MiRNA-24 Downregulates KLF6 Affecting STAT3 Protein Expression and Phosphorylation Regulating Melanogenesis in Cashmere Goat coat.","authors":"Baoyu Zhang, Runlai Liu, Yuxin Zhao, Xinyu Li, Huixian Su, Shufang Li, Jianping Li, Huaizhi Jiang, Qiaoling Zhang","doi":"10.5713/ab.24.0824","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>This study revealed how miRNA-24 downregulates KLF6, regulating melanogenesis in the Cashmere goat coat.</p><p><strong>Methods: </strong>The correlation between miRNA-24 and coat color was determined by Quantitative reverse transcription PCR (RT-qPCR). GO/KEGG analysis and bioinformatics tools were used to screen for target genes. Targeted interactions were confirmed using the dual luciferase assay. The expressions of miRNA-24 and KLF6 on different colored skins of Cashmere goats, and the KLF6 expression in B16-F10 transfected with miRNA-24 mimics/inhibitors/NC were detected by RT-qPCR, Western Blot, and Immunofluorescence. To investigate the pathway that KLF6 influenced, KLF6 was silenced by RNA interference (RNAi), and the melanogenesis pathway's gene expression was examined. The expression and phosphorylation of STAT3 protein in the pathway were inhibited. RNAi and Stattic were utilized to downregulate its protein expression and phosphorylation for detecting tyrosine expression. BALB/c mice were subcutaneously injected with antagomiRNA-24, a miRNA-24 inhibitor. And the influence of miRNA-24 downregulating KLF6 on melanogenesis was examined.</p><p><strong>Results: </strong>MiRNA-24 relates to coat color. KLF6 was chosen as the target gene, and the targeted interaction was confirmed. The expression trends of miRNA-24 and KLF6 were opposing in the skin. The mRNA and protein expression of KLF6 were decreased/increased in the cytoplasmic nucleus of B16-F10 transfected with miRNA-24 mimics/inhibitors. These suggest that KLF6 is downregulated by miRNA-24. When KLF6 was silenced, genes linked to the melanogenesis pathway, including WNT1, WNT2, PRKACA, MAPK1, TYR, and STAT3, were downregulated. When the protein and phosphorylation levels of STAT3 were suppressed, the mRNA level of TYR was decreased, suggesting that KLF6 influences TYR expression by mediating STAT3 expression and phosphorylation. BALB/c mice's skin injected with antagomiRNA-24 showed increased melanin content and decreased KLF6 expression.</p><p><strong>Conclusion: </strong>MiRNA-24 regulates melanogenesis in the Cashmere goat coat by downregulating the KLF6, mediating the STAT3 expression and phosphorylation.</p>","PeriodicalId":7825,"journal":{"name":"Animal Bioscience","volume":" ","pages":""},"PeriodicalIF":2.4000,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Animal Bioscience","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.5713/ab.24.0824","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}
引用次数: 0
Abstract
Objective: This study revealed how miRNA-24 downregulates KLF6, regulating melanogenesis in the Cashmere goat coat.
Methods: The correlation between miRNA-24 and coat color was determined by Quantitative reverse transcription PCR (RT-qPCR). GO/KEGG analysis and bioinformatics tools were used to screen for target genes. Targeted interactions were confirmed using the dual luciferase assay. The expressions of miRNA-24 and KLF6 on different colored skins of Cashmere goats, and the KLF6 expression in B16-F10 transfected with miRNA-24 mimics/inhibitors/NC were detected by RT-qPCR, Western Blot, and Immunofluorescence. To investigate the pathway that KLF6 influenced, KLF6 was silenced by RNA interference (RNAi), and the melanogenesis pathway's gene expression was examined. The expression and phosphorylation of STAT3 protein in the pathway were inhibited. RNAi and Stattic were utilized to downregulate its protein expression and phosphorylation for detecting tyrosine expression. BALB/c mice were subcutaneously injected with antagomiRNA-24, a miRNA-24 inhibitor. And the influence of miRNA-24 downregulating KLF6 on melanogenesis was examined.
Results: MiRNA-24 relates to coat color. KLF6 was chosen as the target gene, and the targeted interaction was confirmed. The expression trends of miRNA-24 and KLF6 were opposing in the skin. The mRNA and protein expression of KLF6 were decreased/increased in the cytoplasmic nucleus of B16-F10 transfected with miRNA-24 mimics/inhibitors. These suggest that KLF6 is downregulated by miRNA-24. When KLF6 was silenced, genes linked to the melanogenesis pathway, including WNT1, WNT2, PRKACA, MAPK1, TYR, and STAT3, were downregulated. When the protein and phosphorylation levels of STAT3 were suppressed, the mRNA level of TYR was decreased, suggesting that KLF6 influences TYR expression by mediating STAT3 expression and phosphorylation. BALB/c mice's skin injected with antagomiRNA-24 showed increased melanin content and decreased KLF6 expression.
Conclusion: MiRNA-24 regulates melanogenesis in the Cashmere goat coat by downregulating the KLF6, mediating the STAT3 expression and phosphorylation.
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