Transcriptomic profiling of Aeromonas hydrophila under nanoplastic and shear stress exposure
For transcriptomic analysis, Aeromonas hydrophila cells from each treatment group were collected from three biological replicates. Cells were harvested by centrifugation, washed, and total RNA was extracted following the Majorbio protocol. After DNase I treatment, RNA purity was assessed using NanoDrop 2000. Strand-specific cDNA libraries were prepared using the TruSeq RNA Sample Prep Kit (Illumina). Briefly, mRNA was fragmented, cDNA synthesized using random hexamers, and dUTP was incorporated during second-strand synthesis. After adapter ligation, uracil-N-glycosylase was used to remove the second strand, followed by PCR amplification. Sequencing was conducted on the Illumina NovaSeq 6000 platform with paired-end reads. Differential expression analysis was performed using DESeq2, and KEGG pathway enrichment was conducted via the Majorbio Cloud Platform.