Fig. 2.

ORF9b protein is not released from 9b-expressing cells. 293T cells were transfected with pCAGGS-3a-HA (3a), pCAGGS (vector), or pCAGGS-9b (9b) independently. At 48 h post transfection, supernatants were clarified, applied to a 20% sucrose cushion, and centrifuged at 100,000 ×g for 3 h at 4 °C. The pellets were suspended in 1×SDS-PAGE loading buffer (Medium). Cell lysates were prepared with 1×SDS-PAGE loading buffer (Cell). Samples were subjected to Western blot analysis with anti-HA antibody to detect 3a protein (37 kDa), anti-9b antibody (11 kDa) to detect 9b protein and anti-actin antibody for internal control.