The mRAN levels of IRAK1 and IRAK4 in active VKH patients, inactive VKH patients and normal controls.

<p><b>A</b>. The mRNA level in PBMCs of IRAK1 of active VKH patients (n = 18), inactive VKH patients (n = 12), and normal subjects (n = 20) was detected by real-time quantitative PCR. <b>B</b>. The mRNA level in PBMCs of IRAK4 of active VKH patients (n = 12), inactive VKH patients (n = 8), and normal subjects (n = 10) was detected by real-time quantitative PCR.</p

The effect of IRAK1/4 inhibitor on the activities of STAT1 and STAT3.

<p>Purified CD4⁺T cells from normal controls (n = 6) were cultured with or without IRAK1/4 inhibitor in the presence of anti-CD3 and anti-CD28 antibodies for 30 minutes. The phosphorylation of STAT1 and STAT3 were analyzed by FCM. <b>A</b>. A representative patient with data near the mean of each group in groups B and C. <b>B</b>. The results represent the percentage of pSTAT1 and pSTAT3 among the CD4⁺T cells and (<b>C</b>) the intensity of pSTAT1 and pSTAT3 in the CD4⁺T cells. Results are expressed as means ± SD.</p

The effect of IRAK1/4 inhibitor on the expansion of Th1 cells and Th17 cells.

<p>Purified CD4⁺T cells from normal controls (n = 8) were cultured with or without IRAK1/4 inhibitor for 3 days in the presence of wither IL-18 or IL-1β. The frequency of Th1 and Th17 cells was analyzed by FCM. <b>A</b>. A representative patient with data near the mean of each group in B and C. <b>B</b>. The results represent the percentages of IFN-γ⁺, IL-17⁺, and IL-17⁺IFN-γ⁺ cells among the CD4⁺T cells in the presence of IL-18. Results are expressed as means ± SD. <b>C</b>. The results represent the percentages of IFN-γ⁺,IL-17⁺, and IL-17⁺IFN-γ⁺ cells among the CD4⁺T cells in the presence of IL-1β. Results are expressed as means ± SD.</p

Effect of IFN-β treatment on IL-9 production.

<p>Splenocytes (A) and DLN cells (B) from the immunized mice following IFN-β or PBS treatment were activated with IRBP_161–180 (20 µg/ml) or anti-CD3/CD28 (1 µg/ml) for 3 days. IL-9 was analyzed by ELISA. Data are representative of three independent experiments with at least five mice per group. ND: not detected.</p

Reduced Production of Higher Alcohols by <i>Saccharomyces cerevisiae</i> in Red Wine Fermentation by Simultaneously Overexpressing <i>BAT1</i> and Deleting <i>BAT2</i>

In red wine, the contents of higher alcohols and ethyl carbamate (EC) are two significant health concerns. To reduce the production of higher alcohols by wine yeast YZ22 with low production of EC, one <i>BAT2</i> was replaced by a <i>BAT1</i> expression cassette first and then another <i>BAT2</i> was deleted to obtain the mutant SYBB3. Real-time quantitative PCR showed that the relative expression level of <i>BAT1</i> in SYBB3 improved 28 times compared with that in YZ22. The yields of isobutanol and 3-methyl-1-butanol produced by mutant SYBB3 reduced by 39.41% and 37.18% compared to those by the original strain YZ22, and the total production of higher alcohols decreased from 463.82 mg/L to 292.83 mg/L in must fermentation of Cabernet Sauvignon. Meanwhile, there were no obvious differences on fermentation characteristics of the mutant and parental strain. This research has suggested an effective strategy for decreasing production of higher alcohols in <i>Saccharomyces cerevisiae</i>

The effect of IRAK1/4 inhibitor on the phosphorylation of NF-κB.

<p>Purified CD4⁺T cells from normal controls (n = 6) were cultured with or without IRAK1/4 inhibitor in the presence of anti-CD3 and anti-CD28 antibodies for 30 minutes. <b>A</b>. A representative example with data near the mean of each group in group B. <b>B</b>. The results represent the mean fluorescence intensity of phosphorylation NF-κB expression in the CD4⁺T cells. Results are expressed as means ± SD.</p

Effect of IFN-β on IL-9 production by polarized Th1, Th17 cells and effctor/memory T cells.

<p>Naïve T cells cultured with or without IFN-β in Th1-polarizing conditions (10 ng/ml IL-12) (A) or Th17-polarizing conditions (20 ng/ml IL-6, 5 ng/ml TGF-β1 and 10 ng/ml IL-23) (B) for 4 days. Effector/memory T cells were cultured with or without IFN-β for 3 days. IL-9 was analyzed by ELISA. Data are representative of three independent experiments.</p

Effect of IFN-β-treated monocytes on the expression of IL-9.

<p>Naïve T cells (A) or effector/memory T cells (B) were cultured with monocyts (Mo) or IFN-β-treated monocytes (IFN-β-Mo) at a ratio of 5∶1 for 3 days. IL-9 levels were detected by ELISA. Data are representative of three independent experiments.</p

Effect of IFN-β on IL-6, TGF-β, IL-23 and IL-10 production by monocytes.

<p>Monocytes from immunized mice on day 13 (or on day 0 and day 16 where indicated), cultured with or without IFN-β (250 U/ml) for 24 hours, were stimulated with LPS (100 ng/ml) for another 24 hours. Cytokines were detected by ELISA. Data are representative of three independent experiments. **p<0.01, NS = not significant.</p

Effect of IFN-β on T cell proliferation.

<p>Naïve T cells (A) or effector/memory T cells (B) from normal mice cultured with anti-CD3 (1 µg/ml) and anti-CD28 (1 µg/ml) in the presence or absence IFN-β for 72 hours. IFN-β-treated monocytes cultured with naïve T cells (C) or effector/memory T cells (D) at a ratio of 1:5 for 72 hours. Proliferation was detected by modified MTT. Data are representative of three independent experiments with at least three mice per group. *p<0.05, NS = not significant.</p