Project description:Background: When pigs underwent apical resection (AR) on postnatal day (P) 1 (ARP1) followed by myocardial infarction (MI) on P28, the hearts had little evidence of scarring; meanwhile, hearts underwent MI on P28 without ARP1 showed large infarcts on P56; and the improvement of ARP1 hearts was driven primarily by cardiomyocyte proliferation. AR and MI were performed ~5 mm (AR) and ~20 mm (MI) above the heart apex; thus, we hypothesize that ARP1 preserved the cardiomyocytes cell-cycle throughout the left ventricle, rather than only near the resection site. Methods: Sections of cardiac tissue were collected from the left ventricle of uninjured pigs and from both the border zone (BZ) of AR and uninjured regions (remote zone, [RZ]) in ARP1 hearts. Cardiomyocyte proliferation was evaluated via immunofluorescence analysis of phosphorylated histone 3 [PH3] and symmetric Aurora B (sAuB). Single nucleus RNA sequencing (snRNAseq) data collected from the hearts of fetal pigs, uninjured pigs, and the BZ and RZ of ARP1 pigs was evaluated via our cell-cycle-specific Autoencoder to identify proliferating cardiomyocytes. Results: Cardiomyocyte PH3 and sAuB expression, and percentage of proliferating cardiomyocytes in snRNA data was significantly more common in both BZ and RZ of ARP1 than uninjured hearts but did not differ significantly between the ARP1-BZ and ARP1-RZ at any time point. Heat shock protein HSPA5 and HSP90B1 were overexpressed at both ARP1-BZ and ARP1-RZ. In AC16 cell, overexpression (and knockdown) of HSPA5-HSP90B1 increased (and decrease) cell-cycle activity. Conclusion: ARP1 preserved proliferative capacity of cardiomyocytes located throughout the left ventricle.
Project description:Co-ip assay was used to conduct protein interaction between ARP1 wild-type cells and ARP1 CBL overexpressed cells. After adhesive strips were cut, samples were sent for mass spectrometry sequencing to search for downstream proteins
Project description:Transcriptome sequencing was used to analyze changes in related signaling pathways and gene expression in ARP1 cells when VCP was knocked down.
Project description:The present study employs Liquid Chromatography-Mass Spectrometry (LC-MS) technology for the identification of proteins, utilizing the Q-Exactive mass spectrometer produced by Thermo Fisher Scientific. Upon the analysis of the ARP1-USP13-OE sample via Proteome Discoverer software, a total of 3631 peptides were matched, corresponding to the identification of 690 distinct proteins. Subsequent analysis of the ARP1-WT sample with the same software revealed 4326 matched peptides, culminating in the identification of 760 proteins.
Project description:To investigated the mechanism of HNRNPA2B1 in MM tumorigenesis, performed m6A IP seq analysis (MeRIP-Seq) to identify the expression changes in stable HNRNPA2B1-knockdown MM cells.60 genes m6A with over 2-fold expression change in shHNRNPA2B compared with control in both ARP1 and H929 cells.44 differentially expressed genes (P<0.05) in both m6A and transcription in shHNRNPA2B compared with control in ARP1 cell.
Project description:To explore the function of PNPO, we constructed ARP1 and H929 pTSB-EV/PNPO-OE cell lines via lentivirus packaging. We found different genes change between EV and OE cells.