Fig. 3: Effects of dimerized HRF/TCTP (d-H/T) and inhibitory effects of dimerized HRF/TCTP-binding peptide (dTBP2) on rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs).

RA-FLSs were treated with vehicle, 75 nM d-H/T, or 75 nM d-H/T plus 75 nM dTBP2 as described in the “Materials and methods”. a Representative microphotographs showing cells that had migrated into the wounded area after 48 h (upper panel, ×50 magnification) and invasive cells that passed through the Matrigel-coated Transwell membrane after 24 h (lower panel, ×100 magnification). b, c Migrated (b) and invading (c) cells were quantified by counting the cells in randomly selected fields. d, e mRNA levels of IL-1β, IL-6, and MMP1 in RA-FLSs and protein levels of IL-6 in culture supernatants were determined at 12 hr by qRT-PCR (d) and at 24 h by ELISA (e) after treatment with vehicle, dTBP2, d-H/T, or d-H/T plus dTBP2. f Change in the proliferation of RA-FLSs treated with vehicle, d-H/T, or d-H/T with dTBP2, as measured by WST-1 assay. g, h Protein levels of ERK, phospho-ERK, and IkB in whole-cell extracts and levels of p65 in nuclear fractions, as determined by western blotting (g); relative density normalized to that of GAPDH or lamin A/C (h). Values are the mean ± SEM of three experiments; *p < 0.05; **p < 0.01; ***p < 0.001 versus vehicle-treated RA-FLSs; # p < 0.05; ### p < 0.001 versus d-H/T-treated RA-FLSs.