Figure 3.

S0859 reduces SLS-dependent NF-κB activation in human keratinocytes. Representative immunofluorescence images (60x) of GAS infections after 4 hours (MOI 10) in the presence of vehicle (DMSO, (A–D) or 100 μM S0859 (E–H). A Rb α NF-κB primary antibody (1:50) and Gt α Rb IgG AlexaFluor 488 (1:200) were used for this experiment. The scale bar indicates 50 µm. Minimal NF-κB nuclear localization was observed in the uninfected + vehicle (A), uninfected + 100 μM S0859 (E), ΔsagA + vehicle (C), and ΔsagA + 100 μM S0859 (G) conditions. Most cells in the WT + vehicle (B) and ΔsagA + sagA + vehicle (D) exhibited strong nuclear localization of NF-κB, which was significantly reduced when these infections were performed in the presence of 100 μM S0859 (F, H). Each panel has an inset that highlights a particular cell, indicating either NF-κB localized to the cytoplasm (A, C, E–H) or the nucleus (B, D). (I) 10 fields from each condition were imaged, and the percentage of cells with NF-κB nuclear localization was calculated by counting the number of cells with NF-κB localized to the nucleus, and dividing by the total number of cells in the field.