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. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: Virology. 2017 Dec;512:113–123. doi: 10.1016/j.virol.2017.09.015

Figure 3.

Figure 3

Δ123 LCLs show an increase in EBNA-LP regulated transcription and protein expression. Transcript expression was tested by treating LCLs with 200 uM 4SU for 1 h. Total RNA was harvested and 4SU incorporated into nascent RNA labeled with biotin and isolated using magnetic streptavidin beads. Nascent and total RNA levels were measured by qPCR and used to calculate (A) relative nascent transcription, (B) relative total transcription, and (C) half-life. GAPDH was used as a normalization control and β-Actin and SETDB1 were used as negative controls. Wp, W promoter driven transcripts. Cp, C promoter driven transcripts, and LMP1 are known to be co-transcriptionally activated by EBNA-LP. *p<0.05, one-sample t-test. Error bars = SD. (D) Western blot of EBNA-LP-regulated viral proteins LMP1 and EBNA2 from WT and Δ123 LCLs. BJAB lysate was used as a negative control. (E) Quantitation of Western blot analysis normalized to WT expression levels. Data represent the average of 6+ donors. *p<0.05, ** p<0.01; one-sample t-test. Error bars = SD.