Fig. 3: NAT10-dependent ac4C modification promotes PAN RNA stability.
A RT-qPCR was performed to detect the mRNA expression levels of PAN, ORF57, ORF65 and ORF-K8.1 in iSLK-KSHV cells with the wild type (NAT10) or the mutant forms (K290A, G641E) of NAT10 overexpression, as well as their control (pCDH). *, P < 0.05, **, P < 0.01, and ***, P < 0.001 by one-way ANOVA versus the pCDH and NAT10 groups, respectively. B RIP assay was performed with anti-Flag antibody, and subsequent RT-qPCR was used to detect PAN RNA enrichment in iSLK-KSHV cells with the wild type (NAT10) or the mutant form (K290A) of NAT10 overexpression and doxycycline induction for 72 h. *, P < 0.05 by two-sided t-test versus the NAT10 group. C RNA decay assays with actinomycin D treatment for 0, 0.5, and 1 h were performed to detect the degradation rate of PAN RNA (Left) and vIL-6 mRNA (Right) in iSLK-KSHV NAT10+/+ (WT) or NAT10+/− (NAT10+/−) cells. *, P < 0.05 by two-way ANOVA versus the WT group. n.s, not significant. D RNA decay assays with actinomycin D treatment for 0, 0.5, and 1 h were performed to detect the degradation rate of PAN RNA in iSLK-KSHV NAT10+/− cells after rescuing WT (NAT10) or mutant (K290A) NAT10. **, P < 0.01 by two-way ANOVA versus the mutant K290A group. Data represent mean ± SEM from n = 3 (C, D) or n = 4 (A, B) biological replicates shown as points. Source data are provided in a Source data file.
