Spc a1

The NSCLC lines (A549, H1975, NCI-H460 and SPC-A-1) and human normal lung epithelial cells (16HBE) were purchased from American Type Culture Collection (ATCC, MD, USA). A549 cells were cultured in DMEM F12 medium (Gibco, NY, USA). H1975, NCI-H460 and SPC-A-1 cells were cultured in RPMI-1640 medium (Gibco). All the cells were maintained in medium supplemented with 10% fetal bovine serum (FBS, Gibco) and cultured at 37°C in an atmosphere with 5% CO₂. miR-187 mimic and negative control (NC) constructs were purchased from GenePharma (Shanghai, China). To assess the effect of miR-187 on cell proliferation, the miR-187 mimic was transfected into A549 and SPC-A-1 cells using Lipofectamine 2000 (Invitrogen, Thermo Fisher Scientific, USA) according to the manufacturer’s protocol.

H1299 and HBE cells were from American Type Culture Collection (Manassas, VA, USA). A549, SPC-A1, PC-9, 95-D, and HEK293T cell lines were from China Academy of Sciences (Shanghai, China).
H1299, 95-D, and HBE cells were maintained in RPMI-1640 medium (Gibco, Gaithersburg, MD, USA) with 10% fetal bovine serum (FBS, HyClone Laboratories, Logan, UT, USA), whereas A549, SPC-A1, PC-9, and HEK293T cells in Dulbecco’s modified Eagle’s medium (Gibco) with 10% FBS in humidified cell incubator, 5% CO₂ at 37 °C.
Cells were transiently transfected with Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) and 200 nM of chemically synthesized miR-411-5p/3p inhibitors or a NC inh and siSPRY4 or siNC (Ribobio, Guangzhou, China) following the standard protocols. qRT-PCR was used to evaluate the RNA level 24 or 48 h after transfection. All the cell phenotypic experiments were performed within 96 h. Relative sequences are listed in Supplementary Table S3.