Fig. 1.

Transfection efficiency 48-hour post-transfection using Lipofectamine 3000 (L3000) varies depending on the airway epithelial cell line used. (A) GFP microscopy of transfected cell cultures 48-hour post-transfection. Images are at 100X magnification and representative of transfections done in duplicate, with the experiment conducted twice. (B) Western blot of whole cell lysates of 1HAE, 16HBE, and NCI-H292 cultures 48-hour post-transfection probed against GFP (26 kDa) and β-actin (42 kDa) as a loading control. Densitometry analysis is summarized, with GFP expression normalized to β-actin. Only comparisons with p > 0.05 are shown. (C) Flow cytometry analysis of 48-hour post-transfection. Transfection efficiency represents the percentage of cells gated positive for GFP, compared to control. Data represents two independent transfection experiments. Only comparisons with p > 0.05 are shown. (D) Alamar Blue cellular viability assay at 24- and 48-hour post-transfection. Fluorescence intensity (560 nm excitation, 590 nm emission) is proportional to the viability of the cell culture. (E) Western blot of whole cell lysates of 1HAE, 16HBE, and NCI-H292 was performed and probed against DNase I (31 kDa) and II (40 kDa). Densitometry analysis is summarized, with expression of each DNase normalized to their respective β-actin. For all experiments, ** = p < 0.01, **** = p < 0.0001. All experiments included two technical replicates and were repeated twice