Quantitative Protein Analysis of AML Cells

The protocol was described in the previous study [13 (link)]. Total protein from AML cells was extracted by Radio-Immunoprecipitation Assay (RIPA) buffer, followed by addition with 1× loading dye and boiling for 10 min. Then, 20 μg protein samples were loaded into 10% dodecyl sulfate,sodium salt-polyacrylamide gel electrophoresis, transferred onto polyvinylidene fluoride membrane, and blocked with 5% defatted milk powder for 1 h at room temperature. After that, the membrane was impregnated in primary and secondary antibodies. Lastly, chemiluminescence was conducted using a highly sensitive electrochemiluminescence solution (Thermo Fisher, USA). The specific primary antibodies used in this study are as follows: anti-Cyclin D1 (ab40754, Abcam), anti-Bcl2 (ab32124, Abcam), anti-Bax (ab32503, Abcam), anti-mTORC2 (ab109081, Abcam), anti-ZFP36 (ab83579, Abcam), anti-Ki-67 (ab92742, Abcam).