Invitrogen rpmi 1640 medium

The A549 NSCLC cell line (cat. no. FH0045) was obtained from the Cell Culture Center of FuHeng Biology, and the A549 cells were maintained in Invitrogen^® RPMI-1640 medium (Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Lonsera; Shanghai Shuangru Biotechnology Co., Ltd.) and penicillin (100 IU/ml)/streptomycin (100 µg/ml) (MedChemExpress). The A549 cell line was authenticated by using short tandem repeat (STR) analysis in combination with sex-typing gene amelogenin detection, and the A549 cell line was compared with the DSMZ STR cell line profile before use. The cells were incubated in a humidified atmosphere containing 5% CO₂ at 37°C.

EJ human bladder carcinoma cells were purchased from ATCC (Manassas, VA, USA) and maintained in Invitrogen RPMI-1640 medium (Thermo Fisher Scientific, Inc., Carlsbad, CA, USA) with 10% fetal bovine serum (Sigma-Aldrich; Merck Millipore, Billerica, MA, USA), 1% non-essential amino acids and 1% penicillin/streptomycin in a humidified incubator at 37°C supplied with 5% CO₂.

The RT4 and T24 human BC cell lines (The Second Affiliated Hospital of Kunming Medical University, Yunnan Institute of Urology, Kunming, China) were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Inc., Waltham, MA, USA), 100 U/ml penicillin and 100 µg/ml streptomycin. Cells were incubated at 37°C in a humidified atmosphere containing 5% CO₂. Invitrogen RPMI-1640 medium, PBS, Opti-MEM I, Lipofectamine 2000 and glutamine were purchased from Thermo Fisher Scientific, Inc.
Two siRNA sequences targeting LASS2 [National Center for Biotechnology Information (NCBI) accession nos. NM013384, NM181747 and NM022075] were synthesized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China): siRNA-1, 5′-GGCUAUUACUUCUUCAAUUTT-3′ and siRNA-2, 5′-CAGUAUUGGUACUACAUGATT-3′. Unspecific control siRNA (si-NC) was also obtained from Guangzhou RiboBio Co., Ltd. The siRNAs were transfected into the RT4 cell line using Lipofectamine 2000, according to the manufacturer's protocol. siRNA was used for the in vitro experiments, and shRNA was used for cell selection and then for the Xenograft model.