Fig 8. SCARB2 is not involved in EV-A71 binding to RD cells, but necessary for viral replication.
(A) EV-A71 binding to RD cells in the presence of anti-SCARB2 pAb. RD cells pretreated with the anti-SCARB2 pAb (50 μg ml-1) were reacted with EV-A71 (4 × 108 genome copies) on ice for 30 min. Then the cells were washed, and cellular and viral nucleotides were extracted. EV-A71 bound to the cell were analyzed by real-time RT-PCR by ΔΔCt method using ATP5F1 mRNA as an endogenous control. As a technical control of detection of reduced copy number, quarter (1 × 108 genome copies) and half (2 × 108 genome copies) amount of EV-A71 was tested in parallel. The relative virus binding of RD cells reacted with 4 × 108 genome copies of EV-A71 without Ab was expressed as 1. (B) EGFP expression in cells infected with EV-A71-EGFP in the presence of anti-SCARB2 pAb at 18 h post-infection. (C) EV-A71 binding to RD and RD-SCARB2-KO clones. EV-A71 bound to the cell were analyzed as in (A). The relative virus binding of RD cells reacted with 4 × 108 genome copies of EV-A71 was expressed as 1. (D) Replication kinetics of EV-A71 in RD and RD-SCARB2-KO clones. Statistical significance was measured for each time point. Results are indicated as the mean and s.e. for three independent experiments (A, B, C) or triplicate analyses (D). Asterisks indicate P < 0.0001.