Fig. 5
From: Optimization of CFTR-mRNA transfection in human nasal epithelial cells
Western blot analyses. Total membrane proteins (40 μg) from HNE cells were isolated using 2 % Triton X-100 and separated in a 7.5 % SDS-PAGE. To identify CFTR, we used a monoclonal anti-CFTR antibody that detects a CFTR band in the range of 170 kDa. In addition, we detected tubulin in the range of 55 kDa for normalization analyses. a HNE cells were transfected with different CFTR-mRNA doses (2.4 μg/cm2, 1.2 μg/cm2, 0.6 μg/cm2). b Statistical evaluation of the relative CFTR protein amount in transfected cells. Transfection with a CFTR-mRNA dose of 0.6 μg/cm2 expressed 1.8-fold more CFTR protein. These results verify the efficiency of the 0.6 μg/cm2 dose as an optimal wtCFTR-mRNA concentration for CFTR expression