human embryonic kidney epithelial cell line hek 293t  (ATCC)

Journal: Cell reports

Article Title: Neutralization escape, infectivity, and membrane fusion of JN.1-derived SARS-CoV-2 SLip, FLiRT, and KP.2 variants

doi: 10.1016/j.celrep.2024.114520

Figure Lengend Snippet: (A) Schematic depicting spike protein mutations that characterize JN.1 and its subvariants. Related XBB.1.5 variants including FLip are included. (B) Variant proportions over time in circulation in the United States (December 2023–May 2024). Data were downloaded from the Centers for Disease Control website and replotted. (C and D) Infectivity in 293T-ACE2 and CaLu-3 cells. Pseudotyped lentiviruses bearing the spike of interest were used to determine entry into (C) 293T-ACE2 and (D) CaLu-3 cells. Relative luminescence readouts were normalized to D614G (D614G = 1.0) for plotting. Bars in (C) and (D) represent means ± standard deviation from six individual measurements of viral infection of different doses ( n = 6). ** p < 0.01, **** p < 0.0001, and ns p > 0.05.

Article Snippet: Cell lines used in this study include human epithelial kidney (HEK) 293T cells (ATCC, RRID: CVCL_1926), 293T cells overexpressing human ACE2 (293T-ACE2) (BEI Resources, RRID: CVCL_A7UK), and human lung epithelial cell line CaLu-3 (ATCC, RRID:CVCL:0609).

Techniques: Variant Assay, Control, Infection, Standard Deviation

Journal: Cell reports

Article Title: Neutralization escape, infectivity, and membrane fusion of JN.1-derived SARS-CoV-2 SLip, FLiRT, and KP.2 variants

doi: 10.1016/j.celrep.2024.114520

Figure Lengend Snippet: (A and B) Representative images of fused cells with 293T cells transfected to produce spike plus GFP, which were co-cultured with (A) 293T-ACE2 cells or (B) CaLu-3 cells. Images were taken 4 h (CaLu-3) or 6.5 h (293T-ACE2) after co-culturing. (C and D) Plots of average area of fused cells for each spike for 3 total replicates ( n = 3) for (C) 293T-ACE2 and (D) CaLu-3 cells. (E and F) Surface expression of spike on 293T cells used to produce pseudotyped lentiviruses was determined using anti-S1 antibody by flow cytometry. (E) Representative histogram depicting relative S1 signal for each variant and (F) a plot of the geometric mean fluorescence values for 3 replicates ( n = 3). (G) 293T cells used to produce pseudotyped lentivirus were lysed and used for western blotting to probe for full length and S2 subunits of spike and GAPDH (loading control). Relative differences between band intensities were determined using NIH ImageJ and normalized to D614G (D614G = 1.0). **** p < 0.0001.

Article Snippet: Cell lines used in this study include human epithelial kidney (HEK) 293T cells (ATCC, RRID: CVCL_1926), 293T cells overexpressing human ACE2 (293T-ACE2) (BEI Resources, RRID: CVCL_A7UK), and human lung epithelial cell line CaLu-3 (ATCC, RRID:CVCL:0609).

Techniques: Transfection, Cell Culture, Expressing, Flow Cytometry, Variant Assay, Fluorescence, Western Blot, Control