Fig. 4.

Acidification in endosomes has a direct impact on TLR3 signaling. RPE1 cells were treated with either (A) ConcanamycinB (ConB) for different time periods, before stimulation with 5 μg/mL poly(I:C); or (B) Dynasore (Dyna) or Dynasore and ConB for 30 min and then exposed to poly(I:C), TNFα, or poly(I:C) complexed with DOTAP (PIC-DOTAP), as indicated. (C) Cells were transfected with TLR3 siRNAs for 3 d before stimulation with poly(I:C) or PIC-DOTAP, as indicated. TLR3 activation in all cases is represented as percentage of cells with NF-κB translocated to the nucleus of at least three independent experiments.