Fig. 2.

The incremental sumoylation level of P inhibits replication of HPIV3. A HeLa cells were transfected with HA-tagged P and increasing amounts of Cer-SUMO1. At 24 h post-transfection, cells were collected and lysed. Samples were further analyzed by SDS-PAGE and immunoblotting with anti-HA monoclonal antibodies and anti-SUMO1 polyclonal antibodies, and with anti-GAPDH monoclonal antibodies to show the loading controls. B vTF7-3-infected HeLa cells were transfected with plasmids encoding N (250 ng), P (62.5 ng), L (100 ng), and the minigenome (50 ng) with increasing amounts of Cer-SUMO1 or Cer-SUMO1-AA (125, 250, 500, and 1000 ng). The relative luciferase expression level of cells transfected with wild-type P was defined as 100% and mock was transfected without P. The expression of P and Cer-SUMO1 or Cer-SUMO1-AA were detected via Western blot analysis with anti-HA and anti-SUMO1 antibodies. GAPDH was used as a loading control. C A549 cells were transfected with plasmids encoding Cer-SUMO1 or Cer-SUMO1-AA. At 24 h post-transfection, the cells were infected with HPIV3 at an MOI of 1. At 48 h post-transfection, the cells were harvested and lysed for Western blot analysis using anti-HN, and anti-GAPDH monoclonal antibodies, and using anti-GFP polyclonal antibodies to detect the expression of Cer-SUMO1 or Cer-SUMO1-AA.***P < 0.001; ns, non-statistically significant.