2.2 Cell culture and transfection
MLE 12 cell line was obtained from Bei Na Chuang Lian Biotechnology
Research Institute (Beijing, China) and followed their instructions to
culture at 37°C. Cell transfection was conducted following the
instruction of Lipofectamine 3000 (Invitrogen, CA, USA). Stably uc.375
or FoxA1 silencing cell lines were screened out as previously reported15. The cells were transfected with 10 pmol/mL of
uc.375 or FoxA1 short-hairpin RNA, or negative control (Sigma-Aldrich)
and selected for 4 weeks with neomycin (1000 μg/ml). To up-regulate
uc.375 or FoxA1, the MLE 12 cells were transfected with uc.375 or FoxA1
overexpression plasmid according to the manufacturer’s protocols.