Figure 2.

CRBN-knockdown THP-1 cells exhibit increases of mROS levels and bactericidal activity. (A,B) Control (Ctrl) and CRBN-knockdown (CRBN^KD) THP-1 cells were treated with or without LPS for different times of 6 and 12 h, stained with MitoSOX-PE, and analyzed by flow cytometry (A). Data are presented as the mean fluorescence intensity (M.F.I) ± SEM from triplicate samples (B). (C) Ctrl and CRBN^KD THP-1 cells were treated with or without LPS for different times, stained with MitoSOX-PE, and analyzed by immunofluorescence microscopy. Data are representative of three independent replicates. (D–F) Ctrl and CRBN^KD THP-1 cells were infected with Salmonella wild type (14028s strain) at a multiplicity of infection of 10 bacteria/cell, as described in the Methods. Cells were lysed with 0.5% deoxycholate in Dulbecco's PBS. Bacteria were diluted (×25), and plated onto LB agar (D). The number of colonies was counted and presented (E). Percentage survival was obtained by dividing the number of bacteria recovered after 0 h (T0), 6 h (T6), 12 h (T12), or 21 h (T21) by the number of bacteria present at time 0 h (T0) and multiplying by 100. All error bars represent mean ±SEM of 3 independent experiments (F) *p < 0.05.