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. 2012 Dec 10;7(12):e51393. doi: 10.1371/journal.pone.0051393

Figure 8. The effect of Rev-dependent export for Let-7 family members and suppressive sequences within HIV-1 in Jurkat cells.

Figure 8

(A) The secondary structure of the miRNA/mRNA duplex of let-7 family members targeting the Bulge sequence was predicted using the RNA-hybrid program. (B) The relative RNA expression levels in the let-7 family members from Jurkat cells were calculated following normalization to the internal control RUN6B. (C) The silencing of RNAs containing the let-7 targeting-sequences in Jurkat cells. (D) The effects of the Bulge and BulgeMut sequences on RNA export by Rev-HA in Jurkat cells. The blue arrow points to the Bulge-containing constructs and the black arrow points to the BulgeMut-containing vectors. Three independent experiments were performed. **P<0.005, *P<0.05. (E) The effect of Rev-mediated export on RNAs in which the suppressive sequences identified in the pol (element 1 in Fig. 5A) and env-nef (element 15 in Fig. 5A) regions were inserted into the Rluc 3′ UTR was assessed in Jurkat cells (black bars, blue arrows). The corresponding vectors with mutations in the repressive sites (lattice and blue bars, black arrows) were also assessed as well (mutation 1-2m-2 in the pol region and 15-3m-2 in the env-nef region; Fig. 5B–G). The bar patterns correspond to the mutational patterns shown in Fig. 5. The Renilla/firefly luciferase value was assessed in each graph. The empty vector C was used as a control, and the results are presented as the mean ± S.D. as a percentage of the control. “pcDNA” denotes the pcDNA3.1(+) plasmid.