3C suppresses PINK1-mediated mitophagy and contributes to Coxsackievirus B3 replication

The data include:Single-cell RNA sequencingMice were randomly assigned to two groups: the Control group (Sham, no viral infection) and the experimental group (VM, Viral Myocarditis, infected with CVB3). At 7 days post CVB3 infection, hearts were collected from five mice per group, and the five hearts in each group were pooled into one composite sample for subsequent experiments. Single-cell sequencing was performed on isolated and sorted heart cells from Balb/c mice.Single-cell suspensions were processed using the 10× Genomics Chromium System (10× Genomics, Pleasanton, CA) to construct 3’ gene expression v3.1 libraries, which were then sequenced on an Illumina Noveseq6000 sequencer. The 10x Genomics platform uses microfluidic technology to encapsulate individual cells and barcoded beads into droplets, forming Single Cell GEMs (Gel Bead-In-EMulsions). Cells in the droplets are lysed to enable mRNA binding to barcoded beads, followed by reverse transcription to generate cDNA libraries. Sample indices were used to trace the origin of each sequence.2.RNA sequencing and AnalysisTotal RNA was extracted from samples using TRIzol reagent (Invitrogen, USA), and RNA integrity was verified via an Agilent Bioanalyzer with a RNA Integrity Number (RIN) ≥8.0. RNA sequencing libraries were constructed using the VAHTS Universal V6 RNA-seq Library Prep Kit (Cat. No. NR604) following the manufacturer’s protocols. Sequencing was performed on an Illumina NovaSeq 6000 platform with paired-end 150 bp (PE150) read length.Differentially expressed genes (DEGs) were identified using the DESeq2 R package. Functional enrichment analyses, including Gene Ontology (GO) and Reactome pathway annotations, were conducted to explore biological functions of DEGs. Gene Set Enrichment Analysis (GSEA) was performed using GSEA software (v4.3.2) with Hallmark gene sets as references, following standard analytical workflows.3.The original image of Western blot.4.The data of qRT-PCR.