Mda mb 415

Primary normal breast epithelial cells (NBECs) were collected from a woman's mammoplasty material at the Department of Plastic Surgery, the First Affiliated Hospital of Sun Yat-sen University (PR China), according to the rules and regulations relating ethical issues on research use of human subjects in China, and established according to the previous report [18 (link)], Breast cancer cell lines, including Bcap-37 was obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China), MCF-7, BT-549, SKBR3, MDA-MB-231,ZR-75-1, MDA-MB-468, MDA-MB-415, MDA-MB-361, ZR-75-30,T47D and MDA-MB-435 were purchased from ATCC and cultured in Dulbecco’s modified Eagle’s medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (HyClone, Logan, UT, USA) and 100 μg/μL streptomycin, and 100 μg/μL penicillin in a humidified incubator containing 5% CO₂ at 37°C.

UACC-812 and MDA-MB-415 cells were purchased from ATCC (Manassas, VA). RS4.11 and HL-60 cell lines were obtained from Ashish Kumar lab (CCHMC). All cell lines were authenticated by STR profiling at Genetica (Burlington, NC). UACC-812 cells were grown in Leibovitz’s L-15 (Gibco) medium with 2 mM l-glutamine containing 20% fetal bovine serum (FBS) and 0.1% antibiotic and antimycotic (Gibco). MDA-MB-415 cells were grown in Leibovitz’s L-15 (Gibco) medium with 2 mM l-glutamine supplemented with 10 μg/ml insulin (Sigma), 10 μg/ml glutathione (Calbiochem), 15% FBS and 0.1% antibiotic and antimycotic (Gibco). SKBR3, BT474, MDA-MB-231, CAL51, T47D cells were cultured in RPMI 1640 (Gibco) containing 10% FBS with 0.1% antibiotic and antimycotic (Gibco). MDA-MB-453 cells were cultured in improved minimum essential medium (Gibco) containing 20% FBS with 0.1% antibiotic and antimycotic (Gibco). All cells were cultured in a humidified atmosphere in 5% CO₂ at 37 °C. All cells were regularly tested for mycoplasma contamination and were negative.

Mouse NIH 3T3 and human breast cancer cell lines MCF-7, MDA-MB-415, and MDA-MB-175 were purchased from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen) supplemented with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin and streptomycin and grown at 37°C under 5% CO₂.

Human breast cancer MDA-MB-415 (ATCC^® HTB-128) [19 (link)], MDA-MB-468 (ATCC^® HTB-132), MCF7 (ATCC^® HTB-22), and MDA-MB-361 (ATCC^® HTB-27) [19 (link)] cell lines were purchased from ATCC (Maryland, USA) and grown in RPMI-1640 medium (Hyclone,Logan, Utah, USA) with 10% fetal bovine serum [FBS] (Gibco, New York, USA) at 37°C in a humidified atmosphere containing 5% CO₂. Cells were transfected with short hairpin RNA (shRNA) plasmids targeting ANKRD22 by Lipofectamine 2000 (11668019, Invitrogen, USA).

A549, BT-20, BT-474, BT-549, CAMA-1, DMS-53, DU4475, HCC38, HCC70, HCC202, HCC1143, HCC1187, HCC1395, HCC1569, HCC1806, HCC1937, HCC1954, HCC2218, HCT-116, Hs578T, Jeko-1, MCF-7, MDA-MB-134-VI, MDA-MB-157, MDA-MB-175-VII, MDA-MB-231, MDA-MB-361, MDA-MB-415, MDA-MB-436, MDA-MB-453, MDA-MB-468, MiaPaCa2, SK-BR-3, NCI-H441, SK-MEL-28, T-47D, U2OS, and ZR-75-1 were obtained from the American Type Culture Collection (ATCC) and cultured according to vendor recommendations. EFM-19 were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ) and SNU-886 were from the Korean Cell Line Bank (KCLB).
Abemaciclib, palbociclib, ribociclib, PIM447, BYL719, LY2090314, everolimus, DYRK1Bi AZ cpd 33 [32 (link)], dinaciclib, GSK2334470, abemaciclib metabolites M2 and M20 [28 (link)], and additional CDK4/6i (see Figure 2C [33 ],) were synthesized by Lilly Research Laboratories. AZD1208 (S7104) and additional palbociclib (S1579, see Supplementary Figure 1A) were purchased from Selleck Chemicals.

Human breast cancer cell lines (MCF-7, SK-BR-3, ZR-75-30, MDA-MB-361, MDA-MB-453, BT474, MDA-MB-435, MDA-MB-415, MDA-MB-231, BT549, ZR-75-1 and Bcap 37) and immortalized normal breast epithelial cells (MCF-10A) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). All cancerous cell lines were grown in Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Invitrogen). Primary normal breast epithelial cells (NBECs) and immortal breast cell line MCF-10A were cultured in keratinocyte serum free medium (KSFM) supplemented with 0.1 ng/mL human recombinant epidermal growth factor and 20 μg/mL bovine pituitary extract (Invitrogen). All cell lines were maintained in a humidified incubator with 5% CO2 at 37°C.