Fig. 2: NSUN2 promotes NPC metastasis through its RNA m5C catalytic ability.
A Gene Set Enrichment Analysis (GSEA) analysis of RNA m5C up-regulated genes revealing upregulated EMT-related pathways in EBV-infected HK-1 cells. B Transwell assays showing the invasion and migration abilities of HK-1-EBV and CNE2-EBV cells with NSUN2 knockdown using shRNA lentivirus while transiently rescuing wide-type NSUN2 (WT), mutant lacking m5C catalytic site Cysteine 271 (C271) and Cysteine 321 (C321) determined as double mutant (DM), and empty vector (EV) as control. Representative images of three biological replicates with similar results were shown. C Lymph node metastasis from mice (n = 7 mice/group) with foot-pad inoculation of CNE2-EBV cells described in B. Extracted lymph nodes were imaged after one month of inoculation. D IHC staining of pan-CK antibodies showing tumor metastasis in lymph nodes displayed in C. Scale bar, 500 μm. Percentages (E) and areas (F) of metastasis in lymph nodes in (C). G Representative images of the lung and liver from mice (n = 5 mice/group) intravenously injected with CNE2-EBV cells described in (B). Hematoxylin and eosin (HE) staining of lung (H) and liver (I) tissue sections from (G), with statistical analysis of metastasis rates presented on the right. Statistical analyses were performed using appropriate tests based on data distribution: the one-way ANOVA test was used for (B, F, H, I). *P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
