Fig 2: Effects of inhibition of interleukin (IL)-2- inducible T-cell kinase (ITK) on the expression of inflammatory cytokines in Graves’ orbitopathy (GO) (n = 4) orbital fibroblasts (OFs).(A, B) Effects of siITK on the expression levels of ITK, IL-17, IL-6, IL-1ß, and tumour necrosis factor-alpha protein, revealed using Western blotting. (*P < 0.05) The results are expressed as the mean ± standard deviation of at least three individual samples, and the graphs are representative of three independent experiments. (C) Quantitative real-time polymerase chain reaction, showed that the mRNA expression of IL-17 was suppressed by siITK treatment. (**P < 0.01) These experiments were performed at least three times using OFs from at least three GO individuals.

Fig 3: Effect of interleukin-2-inducible T-cell kinase (ITK) inhibitors and Bruton’s tyrosine kinase (BTK)/ITK dual inhibitors on the viability and microRNA-155 expression of orbital fibroblasts in Graves’ orbitopathy (GO) (n = 3) and non-GO (n = 3).(A) Based on the MTT results, exposure of cells to = 1 µM of ITK inhibitors and BTK/ITK dual inhibitors for 24 hours did not reduce cell viability to levels below 80% in GO OFs or 90% in non-GO OFs. (B) The expression of miR-155 was significantly downregulated by treatment with ITK inhibitors and BTK/ITK dual inhibitors, in a time-dependent manner in both GO and non-GO OFs. Differences between treated and untreated cells are indicated (in contrast to cells no treated with ITK, BTK/ITK inhibitors, *P < 0.05).