Cell Culture Maintenance Protocol

HeLa PKR-KO cells (provided by A. Geballe) (29 ) were maintained in DMEM supplemented with 5% fetal bovine serum and puromycin (1 μg/ml; Sigma-Aldrich). RK13⁺E3L⁺K3L cells (rabbit) (60 (link)) were maintained in DMEM supplemented with 5% fetal bovine serum, penicillin/streptomycin (100 IU/ml), geneticin (500 μg/ml), and zeocin (300 μg/ml; Gibco). Wild-type (Invitrogen) and PKR-KO T-REx-293 cells were grown in DMEM supplemented with 10% fetal bovine serum, penicillin/streptomycin (100 IU/ml), zeocin (100 μg/ml), and blasticidin (15 μg/ml; Gibco). The T-REx-293 cells stably expressing the bat PKRs were under constant selection with blasticidin (15 μg/ml) and hygromycin (50 μg/ml; Invitrogen).

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Jacquet S., Culbertson M., Zhang C., El Filali A., De La Myre Mory C., Pons J.B., Filippi-Codaccioni O., Lauterbur M.E., Ngoubangoye B., Duhayer J., Verez C., Park C., Dahoui C., Carey C.M., Brennan G., Enard D., Cimarelli A., Rothenburg S., Elde N.C., Pontier D, & Etienne L. (2022). Adaptive duplication and genetic diversification of protein kinase R contribute to the specificity of bat-virus interactions. Science Advances, 8(47), eadd7540.

Publication 2022

puromycin penicillin/streptomycin geneticin zeocin blasticidin hygromycin

Cells Fetal bovine serum Geneticin Hela cells Hygromycin Penicillin Puromycin Rabbit Streptomycin T cells Zeocin

Corresponding Organization : University of California, Davis

Other organizations : University of Arizona, Centre International de Recherches Médicales de Franceville, Northwestern University, Howard Hughes Medical Institute

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Variable analysis

independent variables

Cell lines used: HeLa PKR-KO cells, RK13+E3L+K3L cells, wild-type T-REx-293 cells, and PKR-KO T-REx-293 cells

dependent variables

Not explicitly mentioned

control variables

Media composition: DMEM supplemented with 5% fetal bovine serum, puromycin (1 μg/ml), penicillin/streptomycin (100 IU/ml), geneticin (500 μg/ml), zeocin (300 μg/ml, 100 μg/ml), blasticidin (15 μg/ml), and hygromycin (50 μg/ml)

controls

Positive control: Wild-type T-REx-293 cells
Negative control: PKR-KO T-REx-293 cells

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