human embryonic lung cells (hel299)  (BioResource International Inc)

Journal: PLoS Biology

Article Title: Dual Host-Virus Arms Races Shape an Essential Housekeeping Protein

doi: 10.1371/journal.pbio.1001571

Figure Lengend Snippet: (A) The apical domain of human TfR1 is shown in green, in a co-crystal structure with the Machupo GP1 shown in grey (PDB 3KAS) . The βII-1–βII-2 species-specific virus binding motif (residues 204–212) is highlighted in yellow. The side chains of residue positions identified as evolving under positive selection are shown in red. A human SNP (L212V; rs41301381) has been reported at position 212, shown in blue. (B) MDCK, (C) HEK293, or (D) HEL299 cells were stably transduced to express either 212L TfR1 or 212V TfR1 (or an empty vector) and then infected with various amounts of Machupo pseudovirus. Virus entry is scored by percentage of cells that become GFP positive (+). The error bars in the HEL299 experiment are large due to difficulty in sorting these cells. Nonetheless, this pattern of relative entry between the different TFR1 alleles expressed in HEL299 cells was observed in four independent experiments (not shown). (B, C, D, right-hand panels) Relative cell-surface expression of human TfR1 variants in each cell line was measured on live cells with a fluorescently labeled α-FLAG antibody. In all experiments, three replicates were performed and error bars indicate one standard deviation.

Article Snippet: Human embryonic kidney 293T cells (ATCC CRL-11268), HEK293 cells (ATCC CRL-1573), human embryonic lung HEL299 cells (ATCC CCL-137), and canine kidney MDCK.2 cells (ATCC CRL-2936) were all maintained in Dulbecco modified Eagle's medium (Cellgro) supplemented with 10% fetal bovine serum (Gibco), 100 units ml −1 penicillin, 100 μg ml −1 streptomycin, and 2 mM L-glutamine (Cellgro).

Techniques: Virus, Binding Assay, Residue, Selection, Stable Transfection, Plasmid Preparation, Infection, Expressing, Labeling, Standard Deviation