Figure 3
Inhibition of viral replication by miR-26a in MACR-145 cells was dose dependent and long lasting
.(A-B) MARC-145 cells seeded in 24-well plates were first transfected with 1 μg plasmid (miR-26a or miR-Ctr) or mock transfected, followed by PRRSV VR-2332 infection at the indicated MOI or mock infection at 24 hpt. At 72 hpi, the culture supernatants were measured by TCID50 (A) and the cell lysates were analyzed with an anti-N polyclonal antibody by WB (B). (C-D) MARC-145 cells were first individually transfected with 1 μg plasmid expressing miR-26a or miR-Ctr or mock transfected, followed by PRRSV VR-2332 infection (MOI = 0.1) or mock infection at 24 hpt. At 24, 48, 72, 96 and 120 hpi, the culture supernatants were measured by TCID50 (C) and cell lysates were analyzed by WB (D). (E-F) MARC-145 cells seeded in 24-well plates were first transfected with the miR-26a or miR-Ctr plasmid in increasing amounts (0, 0.25, 0.5, 0.75 and 1 μg), followed by PRRSV VR-2332 infection (MOI = 0.1) at 24 hpt. At 48 hpi, the supernatants of the cell cultures were harvested and measured by TCID50 as the viral titers (E) and the cell lysates were analyzed by WB (F). (G-H) MARC-145 cells were first infected with 0.1 MOI PRRSV VR-2332 and 0, 4, or 8 h later, the cells were transfected with plasmid expressing miR-26a or miR-Ctr or mock transfected. At 48 hpi, the culture supernatants were measured by TCID50 (G) and the cell lysates were analyzed by WB (H). These data are expressed as the mean ± SD and compared to the miR-Ctr group (A, C, G) or “0 μg” group (E) by paired-sampled t-tests; * P < 0.5, ** P < 0.01 and *** P < 0.001.
