NDRG1 is crucial for LANA-mediated DNA replication and episome persistence.

(A) The p8TR plasmid was transfected into BJAB cells along with LANA and NDRG1 or vector plasmids. Cells were collected at 24 hr post-transfection. Western blotting analysis for LANA, NDRG1, PCNA, and Tubulin were performed (left). The amount of p8TR DNA were analyzed by PCR amplification and DNA gel analysis (right). (B) Cells were collected at 72hr post-transfection. Hirt’s DNA from cells were purified and followed by DpnI and ExoIII digestion. LANA-mediated TR DNA replication were determined by qPCR. Data were shown as mean ± SD, n = 3, *p<0.05. (C) Cells were collected at day1,2,3,4 post-transfection. Total TR DNA level were analyzed by qPCR. (D) The p8TR plasmid was transfected into HEK293T-LANA cells along with NDRG1 or vector plasmids. Cells were collected at 24hr post-transfection. Western blotting analysis for LANA, NDRG1, PCNA, and Tubulin were performed (upper). The amount of p8TR DNA were analyzed by PCR amplification and DNA gel analysis (lower). (E) Cells were collected at 72hr post-transfection. Hirt’s DNA from cells were purified and followed by DpnI and ExoIII digestion. LANA-mediated TR DNA replication were determined by qPCR. Data were shown as mean ± SD, n = 3, *p<0.05. (F) Cells were collected at day1,2,3,4 post-transfection. Total TR DNA level were analyzed by qPCR. (G) qPCR analysis assessing the presence of TR DNA in HEK293T-LANA cells with or without NDRG1 after 10 days in the absent of drug selection.