Fig. 2

KMU-11361 regulates IL-2 production and IKK-NF-κB pathway in activated human Jurkat T cells. Jurkat T cells were pretreated with the compound KMU-11361 (0.3,0.6 and 1µM) for 1 h, then subsequently stimulated with anti-CD3/CD28 antibodies for 6 h. A The effect of KMU-11361 on cell proliferation and morphological changes of Jurkat T cells was visualized under a microscope (×100). B Whole-cell lysates were isolated and used to measure the IL-2 protein expression levels by western blotting. C Total RNA was extracted and used to evaluate the mRNA expression levels of pro-inflammatory cytokine (IL2, and TNFa). **p < 0.01, ***p < 0.001. D Jurkat T cells were pretreated with compound KMU-11361 for 1 h, and subsequently stimulated with anti-CD3/CD28 antibodies for 24 h. Measurement of IL-2 concentration in the supernatant obtained from activated T cells was measured using ELISA. E Jurkat T cells pretreated with compound KMU-11361 (0.3,0.6 and 1µM) for 1 h were subsequently stimulated with anti-CD3/CD28 antibodies for 1 h. Whole-cell lysates were isolated and used to measure the protein expression levels of p-IKKα/β, IKKα, p-NF-κB p65, NF-κB p65, p-IκBα, and IκBα by western blotting. F Jurkat T cells pretreated with compound KMU-11361 for 1 h were subsequently stimulated with anti-CD3/CD28 antibodies for the indicated times (15 to 60 min). Whole-cell lysates were isolated and used to measure the protein expression levels of p-IKKα/β, p-NF-κB p65, NF-κB p65, p-IκBα, and IκBα by western blotting. Values in the graph indicate the mean ± SD of three independent experiments (n = 3). The protein expression level of β-actin was used as the loading control