Figure 5

HSP70 inhibits TNF-α induced nuclear translocation of NF-κB in FLSs, as detected using immunocytochemistry. Rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs) were incubated with heat shock protein (HSP)70 (1 μg/ml) for 1 hour, and the FLSs were washed and exposed to tumour necrosis factor (TNF)-α (20 ng/ml) for 30 minutes. The cells were fixed, permeabilized and incubated with rabbit anti-p65 antibody, followed by Cy3-conjugated anti-rabbit immunoglobulin (red). The nuclei of the corresponding cells were demonstrated by Hoechst 33258 staining. Total magnification for images was 200×.