Cell culture
HaCat cells were cultured in Dulbecco’s Modified Eagle Medium (Gibco)
with 10% FBS at 37 degrees Celsius at 5% CO2 and maintained in flat
bottom T75 flasks. SCC9 cells were cultured in a 3:1 mixture of Ham’s
F12 Medium (Gibco) and Dulbecco’s Modified Eagle Medium supplemented
with 10% FBS and maintained at 37 degrees Celsius at 5% CO2 in T75
flasks. For ARO RT-qPCR experiments, cells were grown in 24 well plates
and lipofected with 500 ng of plasmid using lipofectamine 3000
(Thermofisher). In knockdown experiments, two independent siRNAs (Life
Technologies, Silencer Select Pre-Designed siRNA) were co-transfected at
a final concentration of 10 nM during lipofection, and knockdown of the
target gene was verified via RT-qPCR. In experiments involving direct
delivery of single-stranded RNA, cells were grown in 24 well plates and
lipofected using lipofectamine 3000 (Thermofisher) at the concentrations
listed. For Western blot and Boyden chamber experiments, cells were
maintained in 6 well plates prior to seeding.