Fig. 4.
CRX-547 induces less nuclear translocation of NF-κB than CRX-527 in MonoMac6 cells. (A) Gating strategy used for the analysis of NF-κB nuclear translocation (top panels). Live cells were distinguished from dead cells by the presence of a single, well-formed DRAQ5-containing nucleus (Live Cells) after which we gated on live cells (R1) and NF-κB–containing cells (Focused R1) (bottom panels). Nuclear translocation was defined by the colocalization of NF-κB (green) with the nuclear stain DRAQ5 (red), which is indicated by a bright yellow nucleus in the overlaid image of the R1- and NF-κB–containing (Focused R1) cells. (B) Analysis of the nuclear translocation of NF-κB in MonoMac6 cells after incubation with increasing concentrations of CRX-527, CRX-547, vehicle (VEH), or medium for 5, 15, 30, or 120 min. Data are representative of three separate experiments with 3000 to 5000 live focused cells recorded in each experiment.