Figure 4: ATAD3A and HSPD1 are required for virus multiplication.

(A) Knockdown of ATAD3A in BmN-SWU1 cells was confirmed by RT-PCR. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsATAD3A (1 μg), dsATAD3A (2 μg) and dsATAD3A (4 μg). After 48 h.p.t., the transcript levels of ATAD3A were examined by RT-PCR. (B) Effect of dsATAD3A mediated knockdown of BmATAD3A on BmNPV VP39 protein expression. After 48 h.p.t., BmN-SWU1 cells were infected with BmNPV at MOI of 1, and VP39 and Tubulin expression levels was assessed. (C) Effect of dsHSPD1 mediated knock down of BmHSPD1 on HSPD1 transcription level. BmN-SWU1 cells were transfected with dsRNA for dsEGFP (2 μg), dsHSDP1 (1 μg), dsHSPD1 (2 μg) and dsHSDP1 (4 μg). After 48 h.p.t., the transcript levels of HSPD1 were examined by RT-PCR. (D) After 48 h.p.t., the expression levels of VP39 were examined by Western blotting. Tubulin expression levels as control was assessed. (E) Effects of viral DNA replication of knockout ATAD3A on CRISPR/Cas9 system. Transfection with CRISPR/Cas9 system and infected with BmNPV at MOI of 1. (F) Effects of viral DNA replication of knockout HSPD1 on CRISPR/Cas9 system. Transfection with CRISPR/Cas9 system and infected with BmNPV at MOI of 1. At different time-points, total DNA was isolated from knockout cell and quantified by Q-PCR. **P < 0.01.