Fig. 3: Experimental validation of single-cell RNA-seq results.

A UMAP representation of the top 10% of cells expressing NTRK2, with a bar plot quantifying the proportional distribution of cellular states. B FACS detection of TrkB signaling events under different conditions, including an unstained control (unstained), UT, T24, and T48 NPC43 cells. C NTRK2 and SOX2 expressions determined by qRT-PCR. Cells from different conditions were sorted into NTRK2-high and NTRK2-low groups to extract RNA for qRT-PCR. Significant differences (p < 0.05) were observed between T24-high vs. T24-low and T48-high vs. T48-low groups. D Western blot showing SOX2 and TrkB abundance in TrkB-high and TrkB-low cells in UT and T48 conditions. GAPDH was used as a loading control. E qRT-PCR analysis of NTRK2, SOX2, and BZLF1 expression in FACS-sorted TrkB-high and TrkB-low populations (T48 condition). Significant differences (p < 0.05) were detected between T48-high and T48-low groups. F Western blot showing TrkB and Zta abundance in TrkB-high and TrkB-low cells in T48 condition. GAPDH was used as a loading control. G Relative copy numbers of BZLF1, EBER1 (2 EBV genes), and SOX2 determined by qPCR. Cells from different conditions were sorted into TrkB-high and TrkB-low groups to extract DNA for qPCR. H Expressions of NTRK2 and BZLF1 determined by qRT-PCR. Cells were sorted into TrkB-high and TrkB-low groups first (UT-high and UT-low). Then, sorted cells underwent lytic induction treatment for 48 h (UT-high-T48 and UT-low-T48). Significant differences (p < 0.05) were observed between UT-high vs. UT-low and UT-high-T48 vs. UT-low-T48 groups.