Hek 293t

Human HEK-293T cells, the human breast cancer cell lines MDA-MB-231, MDA-MB-468, MDA-MB-436, MCF-7, ZR-75-1, SKBR3 and HS578T, and the human normal mammary epithelial cell line MCF10A were purchased from American Type Culture Collection (ATCC, VA, USA). These cell lines were regularly authenticated by STR analysis and checked for mycoplasma contamination. For the culture of MDA-MB-231, MDA-MB-468, HS578T and HEK-293T cells, high glucose DMEM (Macgene, Beijing, China) supplemented with 10% fetal bovine serum (Gemini, CA, USA), 100 U/ml penicillin (Macgene, Beijing, China) and 100 μg/ml streptomycin (Macgene, Beijing, China) were used. MDA-MB-436, MCF-7, ZR-75-1 and SKBR3 cells were maintained in RPMI-1640 (Macgene, Beijing, China) supplemented with 10% fetal bovine serum (Gemini, CA, USA), 100 U/ml penicillin (Macgene, Beijing, China) and 100 μg/ml streptomycin (Macgene, Beijing, China). MCF-10A cells were cultured as previously described [21 (link)]. All of the cells were incubated at 37 ℃ in a humidified atmosphere with 5% CO2. To inhibit RNA or protein synthesis, cells were treated with actinomycin D (Act D, CST, MA, USA) or cycloheximide (CHX, Selleck, TX, USA), respectively, for the indicated periods of time. MG132 (Selleck, TX, USA) was used to inhibit protein degradation via the proteasome pathway.

Human breast cancer cell lines (MDA-MB-231 and MDA-MB-468), human umbilical vein endothelial cells (HUVECs), THP-1 cells, and HEK293T cells were purchased from American Type Culture Collection (ATCC, Manassas, Virginia, USA). The cells were authenticated by STR analysis (Suzhou, China) and tested for negative mycoplasma contamination using Mycoplasma Detection Kit (Sigma-Aldrich, MP0050, Missouri, USA). MDA-MB-231, MDA-MB-468, HUVEC, and HEK293T cell lines were cultured in DMEM (CM10017, Macgene, Beijing, China), and THP1 cells were maintained with RPMI-1640 medium. The culture medium was supplemented with 10% fetal bovine serum (FBS, Gibco, Carlsbad, California, USA), 100 U/ml penicillin, and 100 µg/ml streptomycin. These cells were incubated in a humidified incubator containing 5% CO₂ at 37 °C.