ACBD3 is required for CVB3 replication. (A) Viability of wild-type (HeLa) and ACBD3 knockout (KO) cells after CVB3 infection. (B) Viral replication assay of wild-type (HeLa) and ACBD3 KO cells using the CVB3 replicon. Transfection with cDNA encoding FLAG-tagged ACBD3 protein into ACBD3 KO cells rescued CVB3 replication, as shown by a viral replication assay using the CVB3 replicon. (C) Wild-type HeLa cells and ACBD3 KO cells transfected with control and plasmid encoding FLAG-tagged ACBD3 protein were infected with CVB3 at an MOI of 5. At 8 h post-infection, cells were fixed and stained with anti-3C antibody (green) and DAPI (blue). (D) Interaction between ACBD3 and the 3A protein of PV1 and CVB3 was confirmed using a mammalian two-hybrid system. (E) A fixed number of HeLa cells and KO cells were plated into six-well plates and incubated. On each day, live cells were counted and normalized to the initial number of cells.
