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MG Milos Gojkovic
PC Pedro P. Cunha
GD Gabriella S. Darmasaputra
LB Laura Barbieri
HR Helene Rundqvist
PV Pedro Veliça
RJ Randall S. Johnson
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All tumor cell lines were cultured in DMEM (11995065, Gibco) supplemented with 10% FBS (10270106, Gibco) and 1% penicillin/streptomycin (10378016, Gibco).

B16-F10-OVA were generated through co-transfection of the transposon vector pT2 containing codon-optimized genes for chicken ovalbumin (OVA; P01012.2), eGFP (ABG78037.1), neomycin phosphotransferase (NeoR; BAD00047.1) and the vector encoding transposase SB11.

OVA, eGFP and NeoR were expressed as a polycistronic peptide interspersed with P2A and furin cleavage sites and synthesized by Gene Art (Thermo Fisher). This was then cloned under the promoter SV40 in the transposon vector pT27BH (gift from Perry Hackett, Addgene plasmid #26556). Plasmid containg the sleeping beauty transposase (pCMV-SB11, Addgene plasmid #26552) was a gift from Perry Hackett.

Transfected cells were cultured with 400 mg/mL G418 (10131027, Gibco) three days post transfection in order to select for transfected cells. Transfection success was confirmed through flowcytometry analysis of eGFP fluorescence. Clonal B16-F10-OVA cell line was then produced through limiting dilution.

Copyright and License information:  ©2021 Gojkovic, Cunha, Darmasaputra, Barbieri, Rundqvist, Veliça and Johnson
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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