Fig. 8.

Elevated levels of G6PD K45 lactylation inhibit cell proliferation in vivo and in vitro (A–C) PHKs, C33A cells, and MEFs were transfected with the indicated plasmids. Cell proliferation was analyzed via cell viability assays. (D) SiHa (HPV16 positive) cells stably expressing shCtrl or shG6PD were further infected with lentiviruses expressing WT G6PD or its mutation, as indicated. (E) G6PD-knockdown cells or those cells rescued by WT G6PD or the K45T or K45A mutation were treated with NAC (2 mM), and cell proliferation was analyzed 5 days after treatment. (F) β-Galactosidase staining was used to detect the level of senescence in MEF cells. Right panel: analysis of β-galactosidase-positive cells. Left panel: representative images. (G–H) Tumors were weighed after mice were euthanized at the endpoint. (I–J) C33A and SiHa cell xenograft tumors were used to determine G6PD enzyme activity. Each dot represents an independent biological replicate in the plots. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared with indicated groups. Statistical significance was determined using unpaired two-tailed t-test. NS, not significant. G6PD, glucose-6-phosphate dehydrogenase; NAC, N-acetyl-l-cysteine.