Transcriptomic analysis was conducted on Siha cells with the HPV 16E7 gene knocked down.

RNA sequencing (RNA-seq) analysis demonstrated that the knockdown of HPV 16E7 in SiHa cells resulted in alterations in gene expression compared to the control group. Total RNA was extracted using Trizol reagent (Invitrogen, CA, USA) following the manufacturer's procedure. The quantity and purity of the total RNA were analyzed using a Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA), with an RNA Integrity Number (RIN) greater than 7.0. Approximately 10 μg of total RNA representing a specific adipose type was used to isolate poly(A) mRNA with poly-T oligo-attached magnetic beads (Invitrogen). Following purification, both poly(A)- and poly(A)+ RNA fractions were fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments were then reverse-transcribed to create the final cDNA library in accordance with the protocol for the mRNA-Seq sample preparation kit (Illumina, San Diego, USA). The average insert size for the paired-end libraries was 300 bp (50 bp). We then performed paired-end sequencing on an Illumina NovaSeq 6000 at lc-bio (China) following the vendor's recommended protocol.