RNA-seq analysis of HGMCs

Illumina TruSeq RNA sample preparation v2 (Illumina, United States) was used to prepare RNA samples for sequencing on a next-generation sequencing platform. Following the manufacturer’s directions, total RNA was extracted from HGMCs using Redzol reagent (SBS Genetech, Beijing, China). FASTQC was used to evaluate read quality, and Seqtk (v1.3-r106) was used to filter it. After that, using HISAT2 with default settings, clean reads were mapped to the human reference genome version 38 (GRCh38/hg38). The expression of transcripts was then determined using the StringTie programme. The mRNAs with |Fold Change| ≥ 2 and p-value < 0.05 were considered significantly differentially expressed (Robinson et al., 2010 (link); Sui et al., 2022 (link)).