Sn12 pm6

Human normal kidney cell line HK-2 and ccRCC cell lines OSRC-2, Caki-1, SN12-PM6, A498, and SW839 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). All the cell lines were frozen in liquid nitrogen after the first 3 passages with 60 ampules of cell stock. After an ampule was thawed, cells were used within 15 passages in every designed experiment. OSRC-2-luciferase was established in previous study [25 (link)]. The cells were cultured in Dulbecco’s Modified Eagle’s Medium (Invitrogen, Grand Island, NY, USA) supplemented with 10% FBS (GIBCO, Brazil), penicillin (25 units/ml), streptomycin (25 g/ml), and 1% L-glutamine at 37 °C with 5% CO2.

293 T cells were purchased from IMMOCELL (catalog number: IM-H222, Xiamen, China). The human kidney proximal tubule epithelial cell line HK-2 and malignant cancer cell lines 786-O, A-498, SN12-PM6, and Caki-1, were purchased from American Type Culture Collection (ATCC, Manassas, VA, United States). The authenticity of these cell lines was confirmed using STR profiling analysis. The cells were cultured in a medium consisting of 90% DMEM (Gibco, catalog number: 11965092, Shanghai, China) supplemented with 10% FBS (Gibco, catalog number: 12483020, Shanghai, China) in an incubator at 37 °C and 5% CO_2.

Five RCC cell lines (ACHN, 786-O, SN12PM6, OS-RC-2 and CAKI-2) and human proximal tubule epithelial cell line HK-2 cell line were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). RCC cells were cultured in RPMI 1640 medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco), 100 units/mL penicillin and 100 μg/mL streptomycin (Sigma, St-Louis, MO, USA). HK-2 cells were maintained in complete RPMI 1640 medium supplemented with keratinocyte serum-free medium, bovine pituitary extract, and human recombinant epidermal growth factor (Invitrogen, Carlsbad, CA, USA).
MiR-212 overexpressing vector, the control vector, miR-212 inhibitor and the negative control were bought from the company of Genecopoeia (Guangzhou, China). Plasmids expressing XIAP were purchased from Addgene (Cambridge, MA, USA). XIAP specific shRNAs were purchased from OriGene (Beijing, China). These vectors were transfected into RCC cells using Lipofectamine 2000 according to the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA).