Figure 6

DHAV-1 3D^pol interacts with the PCBP2 protein and promotes its expression. A, B DEFs were grown in 6‑well plates, and the cells in each well were transfected with 2 µg of pCAGGS‑3D‑HA or 2 µg of pCAGGS‑3 × Flag-PCBP2 alone or co-transfected with 1 µg of pCAGGS‑3D‑HA and 1 µg of pCAGGS‑3 × Flag-PCBP2. The cells were lysed 36 h after transfection, and the lysates were incubated with a mouse anti-Flag or mouse anti-HA antibody and analysed by immunoblotting with an anti-HA or anti-Flag antibody, respectively. C pCAGGS‑3 × Flag‑PCBP2 and pCAGGS‑3D-HA were transfected separately with pCAGGS or pCAGGS‑3 × Flag‑PCBP2 and pCAGGS‑3D-HA were transfected together into cells in 24‑well plates (transfection ratio of 1:1), and cell samples were collected at 24 h after transfection for indirect immunofluorescence experiments to observe the intracellular localization of the 3D^pol and PCBP2 proteins. D DEFs were grown in 12‑well plates and transfected with different doses of 3D‑HA expression plasmids (the amount transfected into each group was 1 µg, and pCAGGS was used to supplement the insufficient group) or with 0.4 µg of pCAGGS‑3 × Flag-PCBP2 plus different doses of pCAGGS‑3D‑HA (the amount transfected into each group was 1 µg, and pCAGGS was used to supplement the insufficient group). Twenty-four hours after transfection, the cell samples were collected, and the protein expression levels of 3D and PCBP2 were measured.