FIG 6.

Silencing of IFITMs suppresses expression of lncRNA IFITM4P. (A) pNL-IFITM1 (0.5 μg) plasmid and plasmid (4.5 μg) carrying shRNA targeting IFITM1, IFITM2, IFITM3, or IFITM4P, were cotransfected into 293T cells. IFITM1 levels were examined by Western blotting 48 h posttransfection. (B and C) In A549 cells infected or not infected with WSN (MOI of 0.5), knockdown efficiency of IFITM1 was determined by Western blotting (B) and qRT-PCR (C). RT-PCR (B) was applied to detect lncRNA IFITM4P expression in IFITM1-silenced and control cells with or without viral infection for 14 h. (D) 293T cells were cotransfected with 0.5 μg pNL-IFITM2 plasmid and 4.5 μg plasmid carrying shRNA targeting IFITM1, IFITM2, IFITM3, or IFITM4P. Western blotting was used to assess the protein levels of IFITM2. (E and F) IFITM4P RNA levels in control and IFITM2 knockdown A549 cells infected or not infected with WSN (MOI of 0.5) for 14 h, were determined by RT-PCR. (G) 293T cells were cotransfected with 0.5 μg pNL-IFITM3 plasmid and 4.5 μg plasmid carrying shRNA targeting IFITM1, IFITM2, IFITM3, or IFITM4P. Western blotting was used to assess the protein levels of IFITM3. (H and I) RT-PCR was used to assess lncRNA IFITM4P levels in IFITM3 knockdown and control A549 cells with or without WSN infection (MOI of 0.5) for 14 h. (J to L) lncRNA IFITM4P levels in control and IFITM1 (J), IFITM2 (K), or IFITM3 (L) knockdown A549 cells infected or not infected with WSN, were determined by qRT-PCR. Data represent mean values ± SEM (n = 3; *P < 0.05; **P < 0.01).