Fig. 2.

HPV16 E6 primarily enhances the PPP and biosynthesis in PHKs and C33A cells. PHKs and C33A cells were stably transduced with a lentiviruses expression vector, HPV16 E6, or HPV16 E7. C33A/PHKs-HPV16 E6, C33A/PHKs -HPV16 E7, and control cells harboring an empty vector were tested for intracellular NADPH (A), NADPH/NADP⁺ (B), GSH (C) GSH/GSSH (D), ROS (E–F), H₂O₂ (G) and protein carbonylation (H). (I–J) Detection of lipid synthesis by performing Nile red staining. Representative images (left) of average fluorescence quantification results (right) are shown. (K–L) EdU was used to determine DNA synthesis in PHKs and C33A cells exogenously expressing HPV16 E6, HPV16 E7, or the vector. ImageJ was used to analyze the proportion of EdU-positive cells. Representative images (left) and quantification results (right) are shown. (M) Cell proliferation rates were determined by performing CCK8 assays. (N) Tumor masses in xenograft nude mice injected with C33A-HPV16 E6 (1 × 10⁶) and C33A-HPV16 E7(5 × 10⁶) cells compared to those in mice injected with the control vector cells. Each dot represents an independent biological replicate in the plots. Data are presented as mean ± SD. Statistical significance was determined using unpaired two-tailed t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared with indicated groups. NS, not significant.