Hgepp

We cultured 4 discrete commercially-available HNSCC cell lines that originated from 4 different sites in the upper aerodigestive tract: [1 (link)] H413 (buccal; Sigma-Aldrich, St. Louis, MO; authenticated November 24, 2006) [14 (link)]; [2 (link)] Detroit 562 (pharynx metastatic to pleura; obtained May 7, 2016 from ATCC, Manassas, VA; authenticated February 17, 2015) [15 (link)]; [3 (link)] FaDu (hypopharynx; obtained May 7, 2016 from ATCC; authenticated February 27, 2014) [16 (link)]; and [4 (link)] Cal 27 (tongue; obtained May 7, 2016 from ATCC; authenticated November 26, 2014) [17 (link)]. ATCC cell lines were authenticated by ATCC through morphological assessment, cytochrome C oxidase subunit I (COI) DNA barcoding, and short tandem repeat (STR) analysis; cells were tested for Mycoplasma contamination using the Hoechst staining and agar culture methods. Sigma-Aldrich obtained the H413 cell line from the European Collection of Authenticated Cell Cultures (ECACC); cells were authenticated using STR analysis and tested for Mycoplasma contamination using the Hoechst staining, agar culture, and Mycoplasma-specific PCR. We additionally obtained primary human gingival epithelial cells that were pooled from 3 healthy female donors (HGEPp; CELLnTEC, Bern, Switzerland) for comparison; cells were tested for contamination using Mycoplasma-specific RT-PCR.

iSLK cells^{54 (link)} were maintained in DMEM medium containing 10% fetal bovine serum (FBS; complete DMEM), 50 μg/ml G418, and 100 μg/ml hygromycin B in the presence of 5% CO₂. iSLK cells containing BAC16 WT or BAC16 mutants were cultured in complete DMEM containing 250 μg/ml G418, and 1000 μg/ml hygromycin and 1 μg/ml puromycin. TREx-(F3H3)-K-Rta BCBL-1 cells that expresses Flag × 3 and HA × 3 tags at the N-terminal region of K-Rta were generated and cultured in complete RPMI 1640 containing 50 μg/ml blasticidin and 100 μg/ml hygromycin B. Vero (ATCC CCL-81) and iVero (dox-inducible K-Rta expression) cells were cultured in the presence of 1 µg/ml puromycin. Human gingival epithelial cells (HGEP), obtained from pooled donors, were purchased from CellnTec (#HGEPp). The cells were cultured in CnT-PRime epithelial cell medium (CellnTec #CnT-PR).