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. 2014 May 1;189(9):1110–1120. doi: 10.1164/rccm.201309-1697OC

Figure 5.

Figure 5.

Association of prostacyclin synthase (PGIS) mRNA and protein expression and promoter variants in human lung tissue quantitative reverse-transcriptase polymerase chain reaction was used to quantitate the PGIS mRNA level in human lung tissue relative to three endogenous control genes. (A) Association of PGIS mRNA levels and the two most common PGIS promoter genotypes in human lung tissue. The common PGIS promoter genotypes were compared; these vary in both variable number of tandem repeats (VNTR) length (4/6 to 6/6) and the presence of single-nucleotide polymorphism (SNP)3 C185T (wt/M1), keeping other SNP compositions constant (T22_G22/4wt_6wt, N = 12; T22_G22/6M1_6wt, N = 15). (B) Association of PGIS protein expression and the two most common PGIS promoter genotypes in human lung tissue. Western blot analysis (15 μg protein extract per lane) from human lung tissue is shown detecting endogenous levels of PGIS protein (50 kD) and two different endogenous control proteins (GAPDH and β-actin). Densitometry of PGIS protein expression relative to GAPDH showed a sixfold increase in PGIS level with the predicted more active PGIS promoter sequence (replicate blots, two-tailed t tests; P = 0.01). The common PGIS promoter genotypes were compared; these vary in both VNTR length (4/6 to 6/6) and the presence of SNP3 C185T (wt/M1), keeping other SNP compositions constant (T22_G22/4wt_6wt, N = 4; T22_G22/6M1_6wt, N = 4). Error bars are the standard error of the mean, and significance (**P = 0.025) was determined by two-tailed t tests.