Cell Culture Protocols for Genetic Manipulation

The 3D4/21 (ATCC, CRL-2843), PK-15 (ATCC, CCL-33), and HeLa (ATCC, CCL-2) cells were cultured in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% FBS (Gibco), 100 units/mL penicillin, and 100 mg/mL streptomycin sulfate (Sangon, Shanghai, China). All cells were grown in monolayers at 37 °C in 5% CO₂. The 3D4/21 P65^−/−, PK-15 ATG5^−/−, and Beclin-1^−/− cells were used and cultivated as previously described [19 (link),20 (link)].

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Wang Q., Zhou L., Wang J., Su D., Li D., Du Y., Yang G., Zhang G, & Chu B. (2022). African Swine Fever Virus K205R Induces ER Stress and Consequently Activates Autophagy and the NF-κB Signaling Pathway. Viruses, 14(2), 394.

Publication 2022

DMEM FBS penicillin streptomycin sulfate

Beclin 1 Cells Hela Penicillin Streptomycin sulfate

Corresponding Organization : Henan Agricultural University

Other organizations : Henan University

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Variable analysis

independent variables

P65 knockout in 3D4/21 cells
ATG5 knockout in PK-15 cells
Beclin-1 knockout in PK-15 cells

dependent variables

Not explicitly mentioned

control variables

Cell culture media (DMEM supplemented with 10% FBS, 100 units/mL penicillin, and 100 mg/mL streptomycin sulfate)
Cell culture conditions (37 °C, 5% CO2)

controls

Wild-type 3D4/21, PK-15, and HeLa cells as positive controls

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