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. Author manuscript; available in PMC: 2017 Mar 9.
Published in final edited form as: Cell Host Microbe. 2016 Mar 9;19(3):409–423. doi: 10.1016/j.chom.2016.02.007

Figure 5.

Figure 5

Pestivirus replication and virus production depend on miRNAs. (A–B) Replication and virus production after infection of MDBK cells with BVDVcp (A) or BVDVncp (B) (MOI=0.1) +/− tinyLNA-17 (0.3µM) (C) Dose-response of BVDVcp infection of MDBK cells (MOI=0.1) to tinyLNA-17. (D) Virus production after electroporation of BVDVcp and m17p3,4 mutant +/− trans-complementation with miR-17p3,4 (50nM). (E) Virus production as in (D) but with trans-complementing the m17p3,4 mutant with miR-17p3,4, miR-17p3,4,15,16 or miR-17p3,4,5,15,16. (F) BVDVcp replication after infection of MDBK cells (MOI=0.1) +/− tinyLNA-let-7 partially inhibiting let-7. (G) Virus production after electroporation of BVDVcp and let-7p3,4 mutant +/− trans-complementation with let-7i–p3,4 (0.1nM). (H) Conservation logo plot showing relative frequency of bases at every position in the 3’ end of BVDV (31 sequences), CSFV (68 sequences) and border disease virus (BDV, 7 sequences) downstream of the AT-rich region. Canonical and non-canonical miR-17 and let-7 sites are indicated. For BVDV, some strains (incl. NADL used in this study) carry a semi-conserved duplication (red line). (I) Virus production after CSFV-luc infection of SK-6 cells (MOI=0.1) +/− tinyLNA-17 or tinyLNA-17p3,4 (0.3µM). (J) Dose-response of CSFV wt or an m17p3,4 mutant 48hrs after transfection of SK-6 cells with miR-17p3,4. (K) Virus production after CSFV wt or m17p3,4 RNA transfection of SK-6 cells transduced with lentiviruses expressing miR-17 or miR-17p3,4 or with the lentivirus backbone (puro, ctr). See also Figure S5–6.