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REH is required for polyadenylation at (pA)d2. (A) LEH and REH mutants of pHBoV1-WH are shown. (B) 3′ RACE. Total RNA was isolated from HEK293T cells transfected with the plasmids shown in panel A, and then 3′ RACE was performed as described in the legend to Fig. 2B. (C) Northern blot analysis. Total RNA isolated from transfected HEK293T cells was resolved in 1.5% agarose gels, transferred to Hybond-N+ membranes, and hybridized with probes spanning nt 4745 to nt 5171.
