Fig 1: LPS stimulation increased MHCII and co-stimulatory molecules expressions, and IL-12p40 secretion. Flow cytometry was used to analyze unstimulated and LPS-stimulated MoDCs for their expression of MHCII, CD80, and CD40 after 6 and 24 h (A) Representative contour plots of MHCII with numbers on plots representing the percentage of MHCII+ cells gated on live cells and the MFI of MHCII+ cells. (B) The upper panel represents the percentage of MHCII+ cells among viable cells. Each circle represents an individual chicken in the corresponding condition. A non-parametric Wilcoxon matched-pairs signed rank test was used to determine statistical differences (ns, not significant). The lower panel represents MHCII MFI determined on viable MHCII+ cells. Each circle represents an individual chicken in the corresponding condition. A two-tailed paired t-test was used to determine statistical differences (****p<0.0001). (C) Representative flow cytometry histograms of CD80 (left panel), and CD40 (right panel) expressions in unstimulated cells (blue), LPS-stimulated MoDCs (orange), and isotype control (grey) after 6 and 24 h Numbers on histograms indicate the MFI values of the corresponding marker. (D) Boxplot of CD80 (upper panel) and CD40 (bottom panel) expression measured as MFI normalized to the isotype control. Each circle represents an individual chicken in the corresponding condition. A two-tailed paired t-test and a non-parametric Wilcoxon matched-pairs signed rank test were used to determine statistical differences (**p<0.01; ***p<0.001). The data from 3 independent experiments are displayed (n=14). (E) The production of IL-12p40 was quantified in the supernatants of unstimulated or LPS-stimulated MoDCs at 6 and 24 h by ELISA. Data are presented as boxplots with individual chicken values represented by circles. A two-tailed paired t-test was used to determine statistical differences (***p<0.001; ****p<0.0001). Data from two independent experiments (n=8) are displayed.
Fig 2: 6 h NDV stimulation enhanced antiviral innate immune response. Chicken MoDC was generated and stimulated with NDV for 6 and 24 h Genes expression was analyzed by RT-PCR. Boxplots representing relative expression of MDA5 (A), LGP2 (B), IFN-α (C), IFN-β (D), TNF-α (E), IL1-β (F), and IL6 (G) determined in unstimulated (blue) and NDV-stimulated MoDCs (purple). Each circle represents an individual chicken in the corresponding condition. The data were normalized to HMBS, TBP, and HPRT1 expressions and calculated according to the 2-ΔΔCT method. A two-tailed paired t-test and a non-parametric Wilcoxon matched-pairs signed rank test were used to determine statistical differences (*p<0.05; **p<0.01). The data from two independent experiments are displayed (n=6). (H) The production of IL-12p40 was quantified in the supernatants of unstimulated or NDV-stimulated MoDCs at 6 and 24 h by ELISA. Each circle represents an individual chicken. A two-tailed paired t-test and a non-parametric Wilcoxon matched-pairs signed rank test were used to determine statistical differences (*p<0.05; ns, not significant). Data from two independent experiments (n=8) are displayed.
Supplier Page from Thermo Fisher Scientific for Chicken IL-12 p40 ELISA Kit