Propagation and Maintenance of Cell Lines

Madin-Darby canine kidney cells (MDCK) (ATCC CCL-34), human lung adenocarcinoma epithelial cells (A549) (ATCC CCL-185), human embryonic kidney HEK293T cells (293T) (ATCC CRL-3216), and Lewis lung carcinoma LL/2 (LLC1) cells (ATCC CRL-1642) of C57BL were purchased. The 293T-IAV-Luc cell line is constructed by our previous study [70 (link)]. The cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS, Lonsera, 711-001S, www.lonsera.cn), 100 U/mL penicillin, and 100 ug/mL streptomycin at 37°C in an atmosphere containing 5% CO₂. The influenza A/WSN/1933 (WSN), influenza A/California/2009/04 (CA04), and influenza A/Puerto Rico/8/1934 (PR8) strains were propagated in MDCK cells. A/Darwin/6/2021/H3N2, A/chicken/Hebei/LC/2008(H9N2) (HB08) strain and Sendai virus (SeV) was propagated in 9-day-old SPF chicken embryonated eggs purchased from Beijing Vital River Animal Technology Co., Ltd. (licensed from Charles River) at 37°C for 72 h.

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Min J., Li Y., Li X., Wang M., Li H., Bi Y., Xu P., Liu W., Ye X, & Li J. (2023). The circRNA circVAMP3 restricts influenza A virus replication by interfering with NP and NS1 proteins. PLOS Pathogens, 19(8), e1011577.

Publication 2023

LLC1) cells DMEM FBS

293t cell Animal Atmosphere Ccl 34 Cells Chicken Eagle Eggs Embryonic Epithelial cells Human Influenza Kidney Lewis lung carcinoma Lung adenocarcinoma Madin darby canine kidney cells Penicillin River Sendai virus Strain Streptomycin

Corresponding Organization : Shenzhen Bay Laboratory

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Variable analysis

independent variables

Cell lines used: Madin-Darby canine kidney cells (MDCK), human lung adenocarcinoma epithelial cells (A549), human embryonic kidney HEK293T cells (293T), and Lewis lung carcinoma LL/2 (LLC1) cells
Influenza A virus strains used: A/WSN/1933 (WSN), A/California/2009/04 (CA04), A/Puerto Rico/8/1934 (PR8), A/Darwin/6/2021/H3N2, A/chicken/Hebei/LC/2008(H9N2) (HB08), and Sendai virus (SeV)

dependent variables

Cell maintenance conditions: Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 U/mL penicillin, and 100 ug/mL streptomycin at 37°C in 5% CO2 atmosphere

control variables

The 293T-IAV-Luc cell line constructed in a previous study [70]
SPF chicken embryonated eggs purchased from Beijing Vital River Animal Technology Co., Ltd. (licensed from Charles River) used for virus propagation

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