Fig. 5.

The effects of knockdown expression of Na⁺/K⁺-ATPase α1, PI3K, or PDK1 on the TGEV activity. A. Knockdown of Na⁺/K⁺-ATPase α1 expression activated PI3K_PDK1_RSK2 axis pathway and suppressed TGEV activity. B & C. Knockdown of PI3K (B) or PDK1 expression (C) reversed the ouabain-inhibited TGEV activity/TGEV N protein expression. The validated knocked down cells were seeded onto a 6 well-plate, 1.8 × 10⁶ cells/well, and cultured for 24 h prior to harvesting for western analysis with the antibodies indicated. NS: no significance; **, P < .01. Na⁺/K⁺-ATPase α1, p-PDK1, PI3Kα and N protein were normalized with GAPDH. p-RSK2 was normalized with regular RSK2. Results shown are represented results or averages ± SD from three independent experiments. The gene names for Na⁺/K⁺-ATPase α1, PDK1 and PI3Kα respectively are ATP1A1, PDPK1 and PIK3CA.