FIG 4.

T4519 regulates cytokine/chemokine responses. (A and B) PMA-differentiated THP-1 cells were infected with the indicated bacterial strains and were cultured as described for Fig. 1. (A) Reverse transcription-qPCR shows GAPDH-normalized expression of cytokines and chemokines at 12 h postinfection. Error bars, means ± standard deviations for three independent experiments. Asterisks indicate significant differences from results for Ty2 infection by Student's t test (*, P < 0.05; **, P < 0.01; ***, P < 0.001). (B) Cytokine and chemokine levels in culture supernatants after 24 h of infection were determined by ELISA. (C) A serum ELISA was performed 48 h after infection of mice. (D) PMA-differentiated THP-1 cells were infected with strain Ty2 or Ty2Δt4519 and were cultured as described for Fig. 1 for the indicated times. (Top) Immunoblots of total-cell lysates probed for IκB-α (NF-κB activation) and for phosphorylated (indicating activation) and total (loading control) MAP kinase (ERK) and AKT. Tubulin was used as a loading control for IκB-α. (Bottom) Immunoblots of nuclear extracts from infected cells were probed for p65 (NF-κB activation) and histone 3 (loading control).