IMMT Knockdown in MDA-MB-231 Cells

The cell culture and transfection were conducted as previously described [31 (link)]. Briefly, MDA-MB-231 cells were grown in DMEM (HyClone) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific). The existence of mycoplasma was determined using the e-Myco™ plus Mycoplasma PCR Detection Kit (iNtODEWORLD, MA, USA). For the knockdown of IMMT, 20 nM of IMMT-targeting siRNA (4392420, Thermo Fisher Scientific) and its corresponding negative control RNA (4390843, Thermo Fisher Scientific) were introduced into cells using the Lipofectamine™ RNAiMAX Transfection Reagent (LMRNA015, Invitrogen, Carlsbad, CA, USA). The siRNA transfection efficiency was determined by western blot 72 h after transfection.