Figure 3: ACBD3 is necessary for EV71 replication.
From: The Golgi protein ACBD3 facilitates Enterovirus 71 replication by interacting with 3A
(a) RD cells were transfected with siRNA against ACBD3. At 48 h, cells were harvested for Western blot analysis. (b) RD cells were treated as indicated in (a). Cell viability was detected by using Cytotoxicity Detection KitPlus (LDH). (c) RD cells were transfected with siRNA against ACBD3. After 48 h, cells were infected with EV71 at MOI of 1 PFU/cell. At 24 h, total RNA was extracted and the viral RNA levels of EV71 were evaluated by quantitative real-time PCR using SYBR Green. Data are expressed as fold change of the EV71 RNA level relative to the control using the ΔΔCT method as described in the Materials and Methods. (d) RD cells were treated as indicated in (c). Cells were harvested for tissue culture infective dose (TCID50) assay. (e) Plaque assay of viruses were performed on RD cells at 37 °C. Monolayers were stained with crystal violet at 72 h post infection. Data shown are representative of three independent experiments. (f and g) RD cells were transfected with siRNA against ACBD3. At 48 h, cells were infected with different EV71 strains as indicated. After 12 (f) or 24 (g) h, total RNA was extracted and the viral RNA levels of EV71 were evaluated by quantitative real-time PCR using SYBR Green.
