Figure 2
RT-qPCR analysis of linear LMP-2 and circLMP-2_e5 in various EBV-positive cell lines. Expression of (a) circLMP-2_e5, and linear (b) LMP-2A and (c) LMP-2B in EBV-positive cell lines with different latency programs in latent and lytic states. Akata was reactivated using Goat (Fab)2 fragment anti-human immunoglobulin G (IgG) for 24 h while C17 was reactivated by transfection with BZLF1-expressing plasmid p509 for 72 h using X-tremeGene HP DNA transfection reagent. The remaining cell lines were induced into lytic cycle using TPA and SB for 72 h except for NPC43 which was induced for 48 h. C666-1 cells were unable to be reactivated and this is consistent with previous reports of the abortive nature of the lytic reactivation in these cells. Human embryonic kidney cell line (HEK293T), an EBV-negative cell line was used as a negative control. Data was normalized to UBC and relative to gene expression in Akata cells. Data represents the mean ± SD of two independent experiments. (d, e) Expression of circLMP-2_e5 and linear LMP-2A in newly generated LCLs. Data was normalized to UBC and relative to gene expression in C666-1 cells. Data represents the mean ± SD of two independent experiments. ND not-detected.
