Skip to main content
. 2019 Oct 9;10(10):783. doi: 10.3390/genes10100783

Figure 3.

Figure 3

Expression analyses of selected genes in human PBMCs and lung fibroblast cells and serum IL-1β concentration. (A) Real-time PCR analysis of NLRP1, NLRP4 and NLRP8 mRNA was performed on cDNA samples from COPD patients (N = 4) and WI38 cell line (human lung fibroblasts). Total RNA was isolated either from peripheral blood mononuclear cells (PBMC) or cell line, and reverse-transcribed to cDNA. GAPDH was used as a reference or endogenous control. For quantification see Materials and Methods. (B) Real-time PCR analysis of NLRP1 mRNA expression levels. The expression analysis was performed on 15 cDNA samples from heathy individuals and equal number of COPD patients (RNA was isolated from PBMC). GAPDH was included as a reference or endogenous control. ∆CT values of NLRP1 were normalized to GAPDH CT values and ∆CT is shown on the graph. Each column represents the average value of ∆CT of 5 individuals with the given genotype (A/A, A/T or T/T). The experiment was conducted two times in duplicates. (C) Concentration of the serum IL-1β in healthy and COPD individuals. IL-1β concentration was determined in the plasma samples separated from the healthy donors (N76) and the individuals with diagnosed COPD (N = 100) (NLRP1 genotype (rs12150220) was known for each sample). Analysis was performed with a ProcartaPlex High Sensitivity Assay, according to manufactures protocol, which can be found in the Materials and Methods section. (D) The concentration of serum IL-1β is related to rs12150220. The relationship between IL-1β concentration and NLRP1 (rs12150220) genotypes AA (N = 24) vs. AT+TT (N = 76) in the COPD serum samples. AT and TT genotypes carriers had higher IL-1β levels compared to the AA genotype carriers.