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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 5.

Figure 5

pre-miR-BHRF1-2/2* and 1-3 miRNA excision regulates protein expression in cis. (A) The BHRF1 3′UTR, containing the pri-miR-BHRF1-2 and 1-3 stem-loops (Fig. 4), was cloned from both the WT and Δ123 EBV virus into the 3′UTR of a FLuc expression vector. These vectors, along with a NanoLuc internal control vector, were transfected into both 293T cells and RNaseIII−/− cells and luciferase activity measured. Expression level was normalized to NanoLuc followed by an empty FLuc vector control. Relative 293T luciferase expression was normalized to the expression detected in the RNaseIII−/− cells to observe differences due to Drosha processing. Five biological replicates were performed. **p<0.01, one-sample t-test, and student’s t-test. Error bars = SD. (B) Northern blots show miRNA expression from the BHRF1 3′UTR firefly luciferase vector. Blots were probed with a U6-specific probe to demonstrate equal loading. * Indicates pre- and mature miRNAs.