DataSheet_1_Histone lysine-specific demethylase 1 regulates the proliferation of hemocytes in the oyster Crassostrea gigas.docx

• Main Authors: Xiaoyu Gu (576058), Xue Qiao (1409857), Simiao Yu (4577854), Xiaorui Song (423861), Lingling Wang (66269), Linsheng Song (154137)
• Format: Dataset
• Language: unknown
• Published: 2022
• Subjects: Immunology; Applied Immunology (incl. Antibody Engineering; Xenotransplantation and T-cell Therapies); Autoimmunity; Cellular Immunology; Humoural Immunology and Immunochemistry; Immunogenetics (incl. Genetic Immunology); Innate Immunity; Transplantation Immunology; Tumour Immunology; Immunology not elsewhere classified; Genetic Immunology; Animal Immunology; Veterinary Immunology; Crassostrea gigas; hemocytes; LSD1; histone demethylation; cell proliferation; Pacific; Pacific oyster
• Online Access: https://doi.org/10.3389/fimmu.2022.1088149.s001

Background Lysine-specific demethylase 1 (LSD1) is an essential epigenetic regulator of hematopoietic differentiation, which can specifically mono-methylate H3K4 (H3K4me1) and di-methylate H3K4 (H3K4me2) as a transcriptional corepressor. Previous reports have been suggested that it participated in hematopoiesis and embryonic development process. Here, a conserved LSD1 (CgLSD1) with a SWIRM domain and an amino oxidase (AO) domain was identified from the Pacific oyster Crassostrea gigas. Methods We conducted a comprehensive analysis by various means to verify the function of CgLSD1 in hematopoietic process, including quantitative real-time PCR (qRT-PCR) analysis, western blot analysis, immunofluorescence assay, RNA interference (RNAi) and flow cytometry. Results The qRT-PCR analysis revealed that the transcripts of CgLSD1 were widely expressed in oyster tissues with the highest level in the mantle. And the transcripts of CgLSD1 were ubiquitously expressed during larval development with the highest expression level at the early D-veliger larvae stage. In hemocytes after Vibrio splendidus stimulation, the transcripts of CgLSD1 were significantly downregulated at 3, 6, 24, and 48 h with the lowest level at 3 h compared to that in the Seawater group (SW group). Immunocytochemical analysis showed that CgLSD1 was mainly distributed in the nucleus of hemocytes. After the CgLSD1 was knocked down by RNAi, the H3K4me1 and H3K4me2 methylation level significantly increased in hemocyte protein. Besides, the percentage of hemocytes with EdU-positive signals in the total circulating hemocytes significantly increased after V. splendidus stimulation. After RNAi of CgLSD1, the expression of potential granulocyte markers CgSOX11 and CgAATase as well as oyster cytokine-like factor CgAstakine were increased significantly in mRNA level, while the transcripts of potential agranulocyte marker CgCD9 was decreased significantly after V. splendidus stimulation. Conclusion The above results demonstrated that CgLSD1 was a conserved member of ...