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.