Fig. 5

TAK1 inhibition with takinib alters the phosphorylation of kinases and NF-κB protein expression levels in RA-FLS cells. Representative phospho-kinase blots from RA-FLS cells treated with takinib or DMSO and stimulated with TNF (30 ng/mL) for 30 min (a). Relative phosphorylation of p38α, JNK1/2/3, Akt 1/2/3, STAT3, PYK2, and Fgr from naïve (un-stimulated n = 4 ± SEM, vehicle (TNF + DMSO) n = 4 ± SEM, and takinib (TNF + takinib 10 μM) n = 4 ± SEM (b–g) (two-way ANOVA, Dunnett’s post hoc). Volcano plot of NF-κB-associated protein expression 24 h post-TNF (30 ng/mL) stimulation (h). Expression of BCL10, CARD6, IkBε, IKK1, IKK2, IL-1R, Myd88, NF-κB1, STAT2, and TNF RI from vehicle- and takinib-treated RA-FLS cells (i) vehicle (DMSO) control (n = 4 ± SEM), 10 μM takinib (n = 4 ± SEM), (Students t test). *p < 0.05, **p < 0.01, ***p < 0.001. RFU = relative fluorescent units