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. 2020 Jul 14;12(7):756. doi: 10.3390/v12070756

Figure 5.

Figure 5

The coiled-coiled domain (CCD) and evolutionary conserved domain (ECD) of Beclin1 were essential for the facilitation of EV71 replication. (A) Schematic diagram of Beclin1 and its mutants. (B) HEK293T cells were co-transfected with plasmids encoding GFP-3D and Flag-vector as a negative control: Flag-Beclin1, Flag-Beclin1 BD, Flag-Beclin1 BD+CCD, Flag-Beclin1 BD+CCD+ECD, Flag-Beclin1 CCD+ECD, Flag-Beclin1 ECD. The cell lysates were immunoprecipitated with an anti-Flag antibody and then immunoblotted with the indicated antibodies. (C and D) RD cells were transfected with plasmids expressing Flag-vector, Flag-Beclin1 and various mutants for 24 h and infected with EV71 (MOI = 2) for another 12 h. The expression RNA level of EV71-VP1 was detected by Real-time PCR with GAPDH as a control (C). The protein level of EV71-3D was detected by Western blot (D). Graph expressed as mean ± SD, ns, not-significant, ** p < 0.01.