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Figure 1 | Respiratory Research

Figure 1

From: The role of bronchial epithelial cells in the pathogenesis of COPD in Z-alpha-1 antitrypsin deficiency

Figure 1

Expression of M- or Z-AAT in the 16HBE cell line. ( A ) 16HBE cells expressing M- or Z-AAT were assayed for AAT expression by SDS PAGE and Western blot analysis. 24 hours after transfection, proteins from 50 μl of cell culture media or from 10 μg or 10 μl of NP40-soluble and insoluble cell lysates respectively were separated by SDS-PAGE, transferred to PVDF membranes, and hybridized with anti-AAT antibody to detect total AAT protein. Purified human serum AAT (hAAT) was used as positive control. Typical fluorograms from three independent experiments, which gave superimposable results, are shown. ( B ) Nondenaturing PAGE and Western blot analysis using anti-AAT antibody to detect total AAT in 10 μg of NP40-soluble cell lysate and 10 μl of cell culture media from 16HBE cells expressing M- or Z-AAT. The migration of polymeric and monomeric AAT species is indicated. Typical fluorograms from three independent experiments, which gave superimposable results, are shown. ( C ) Nondenaturing PAGE and Western blot analysis using the antipolymeric Z-AAT antibody ATZ11 in NP40-soluble cell lysate fraction (10 μg) and ( D ) cell culture media (50 μl)from 16HBE cells expressing M- or Z-AAT. Typical fluorograms from three independent experiments, which gave superimposable results, are shown. Definition of abbreviations: AAT = alpha-1 antitrypsin; h-AAT = human serum AAT; PAGE = polyacrylamide gel electrophoresis; PVDF = polyvinylidene fluoride; SDS-PAGE = sodium dodecyl sulphate polyacrylamide gel electrophoresis.

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