Fig. 2.
Analysis of PBEF gene expression and protein levels in canine BAL/serum, and murine BAL in ALI. The left panel demonstrates that in each ALI model (canine or murine) as well as in BAL cells RNA from humans with ALI, that PBEF mRNA levels were validated by semi-quantitative duplex RT-PCR. Statistical comparative analysis of integrated density value (mean ± SEM) between control and ALI groups was performed using the paired t-test and unpaired t-test. The right panel reflects Western blot results that confirm PBEF protein content in lung tissue, BAL or serum. For BAL protein determinations, total protein (10 μg) of each sample was separated by 12% SDS-PAGE and immunodetected by western blot using the anti-canine PBEF antibody. Statistical comparative analysis of integrated density value (mean ± SEM) between control and ALI groups was performed using the unpaired t-test. PBEF protein levels in human ALI BAL and serum were significantly increased relative to healthy controls. These results support PBEF as a potential biomarker in ALI and further validate the microarray-based enhanced PBEF expression in animal and human ALI.