Figure 3.4 Pro-inflammatory cytokine and chemokine profile in gastric environment. (A-C) Semi-quantitative SYBR-Green real time PCR for CCL20, hBD-2 and IL-23p19 mRNA respectively. Gene expression was normalized to the housekeeping gene β-actin and arbitrary values were further normalized to group N. (D) Semi-quantitative TaqMan Gene expression assay of IL-1β mRNA. Gene expression was normalized to the housekeeping gene GAPDH and arbitrary values were further normalized to group N. (E-G) Milliplex xMAP assay for ex vivo IL-6, TNF-α and IL-1β concentrations. (H) ELISA for ex vivo hBD-2 concentrations. (I-L) Subclassification of group P individuals into PC+ or PC- groups for analysis of ex vivo concentrations of IL-6, TNF-α, IL-1β and hBD-2 respectively. 63 Figure 3.5 Other pro-inflammatory cytokines and chemokines analysed. Milliplex xMAP assay for ex vivo IL-1α, IL-1Ra, IL-2, IL-8, IL-10, IL-12p40, IL-15, MCP1 and RANTES concentrations. 64 . Figure 3.4 Pro-inflammatory cytokine and chemokine profile in gastric environment. (A-C) Semi-quantitative SYBR-Green real time PCR for CCL20, hBD -2 and IL -23 p19 mRNA respectively. Gene expression. normalized to the housekeeping gene β-actin and arbitrary values were further normalized to group N. (D) Semi-quantitative TaqMan Gene expression assay of IL-1β mRNA. Gene expression was normalized to. vivo hBD -2 concentrations. (I-L) Subclassification of group P individuals into PC+ or PC- groups for analysis of ex vivo concentrations of IL-6, TNF-α, IL-1β and hBD -2 respectively. 64