Mda mb 468

Human TNBC cell lines MDA-MB-231 (p53R280K) and MDA-MB-468 (p53R273H) were purchased from Cell Bank in Chinese Academy of Sciences in Shanghai. MDA-MB-231 was maintained in DMEM, whereas MDA-MB-468 Cells were cultured in Leibovitz’s L-15 supplemented with 10% FBS (HyClone; GE Healthcare Life Sciences, Logan, UT, USA), 100 U/mL penicillin and 100 μg/mL streptomycin at 37 °C with 5% CO₂. When growth reached 80% confluence, MDA-MB-231 and MDA-MB-468 were treated in triplicate with 2 μM and 0.2 μM BEZ235, respectively, with 0.01% DMSO control^{20 (link)}, which were incubated for 48 h and 72 h, respectively. The cells were removed as raw materials for subsequent RNA analysis.

Human TNBC cell lines MDA‐MB‐231 (p53R280K), MDA‐MB‐468 (p53R273H) and MDA‐MB‐157 (p53 null) were obtained from the Department of Laboratory Medicine, Chongqing Medical University (China, Chongqing). MDA‐MB‐231 was maintained in Dulbecco's modified Eagle's medium (DMEM), whereas MDA‐MB‐468 and MDA‐MB‐157 were grown in Leibovitz's L‐15 (L15) supplemented with 10% fetal bovine serum (HyClone; GE Healthcare Life Sciences, Logan, UT, USA), penicillin (100 U·mL⁻¹) and streptomycin (100 μg·mL⁻¹; Sigma‐Aldrich; Merck KGaA, Darmstadt, Germany). Cells were incubated in a humidified incubator under 5% CO₂ at 37 °C. NVP‐BEZ235 was purchased from LC Laboratories (Woburn, MA, USA), and MG‐132 and 3‐methyladenine (3‐MA) were purchased from Sigma‐Aldrich (Merck KGaA). All primary antibodies, including P53 (#2524), LC3 (#4108), phosphorylated (p)‐Akt (#9271), p‐mTOR (#2971), p‐P70S6K (#9205), p‐AMP‐activated protein kinase (p‐AMPK; #2535), p‐MDM2 (#3521), Akt (#4685), mTOR (#2983), P70S6K (#2708), p‐AMPK (#5832), p‐MDM2 (#86934) and glyceraldehyde‐3 phosphate dehydrogenase (GAPDH; #2118) were obtained from Cell Signaling Technology (Beverly, MA, USA).