Fig. 2
From: Optimization of chemical transfection in airway epithelial cell lines
Transgene expression and cellular viability vary between transfection reagents used. (A) GFP fluorescence microscopy of 1HAE, 16HBE, and NCI-H292 cell cultures 48-hour post-transfection with Lipofectamine 3000 (L3000), FuGENE HD (FuGENE), ViaFect, jetOPTIMUS, EndoFectin, and calcium phosphate (CaP). 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. (C) Flow cytometry analysis of 1HAE, 16HBE, and NCI-H292 cultures 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, with pairwise comparisons made between reagents that yielded significantly different transfection efficiencies. (D) Alamar Blue cellular viability assay on 1HAE, 16HBE, and NCI-H292 cells 24- and 48-hour post-transfection with the tested reagents normalized to either 24- or 48-hour control. For all experiments, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001. All experiments included two technical replicates and were repeated twice