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. 2020 Nov 23;94(24):e01551-20. doi: 10.1128/JVI.01551-20

FIG 7.

FIG 7

The DENV NS2B3 protease complex associates with Nrf2. (a) 293T cells were transfected with human Nrf2-FLAG (2 μg), NS2B3-WT-V5 (1 μg), NS2B3-S135A-V5 (1 μg), or an empty vector, alone or in combination. After 16 h, cells were collected and lysed. (Top) Immunoprecipitation (IP) with anti-FLAG was performed, and the amount of coprecipitated NS2B3 (immunoblot [IB]: anti-V5) (upper panel) or Nrf2 (IB: anti-Nrf2) (lower panel) was analyzed by Western blotting. (Bottom) Total lysates were also analyzed for the expression of Nrf2-FLAG by using an anti-FLAG Ab and for the expression of NS2B3-V5 (WT or S135A) by using an anti-V5 Ab. GAPDH was used as a loading control. (b) A549 cells were infected with DENV (MOI, 1), collected after 24 h and 48 h of infection, and lysed. (Top) Immunoprecipitations with anti-Nrf2 or an isotype-matched control IgG Ab were performed, and the amount of coprecipitated DENV NS3 (upper panel) or Nrf2 (lower panel) was analyzed by Western blotting. (Bottom) Total lysates were also analyzed for the expression of Nrf2 and DENV NS3. β-Actin was used as a loading control. Results are representative of two independent experiments.