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. 2017 Mar 23;8(27):43822–43837. doi: 10.18632/oncotarget.16517

Figure 3. LeRNA inhibits RABV replication by interfering the binding of RABV nucleoprotein to genomic RNA.

Figure 3

(A) SK-N-SH cells were infected with rAAV-U6-leRNA-64 nt or rAAV-U6-control RNA AAV virus and after 24 h, infected with DRV-AH08 (MOI is 0.01). The levels of DRV-AH08 viral genomic RNA (vRNA) were detected by Real Time PCR at 12 h post-infection with DRV-AH08. (B) Schematic diagram of the precipitation (IP) of nucleoprotein to genomic RNA under leRNA and control RNA overexpression respectively. (C) N protein with Fc, P protein and DRV-AH08 genome fragment were co-transfected with leRNA or control RNA in HEK-293T cells, the level of N protein captured genomic RNA was determined by Real Time PCR after 48 h transfection. The ratio of precipitated RNA/Input RNA of leRNA group has been normalized to the ratio of control RNA group. (D) The input of N protein in precipitation assay were detected by Western blot. The ratio of signal intensity of input N protein was quantified using ImageJ. (E) The input of viral genomic RNA in precipitation assay were detected by RT-PCR. The ratio of signal intensity of input genomic RNA were quantified using ImageJ. All experiments were repeated at least six times. All values are the mean ± SEM. P values were determined by Student's t-test. *P < 0.05; **P < 0.01; ***P < 0.001.