Fig. 2.

AMPKα isoforms expression in TNBC lung colonization. (A) MDA-MB-231 and MDA-MB-468 cells were transfected with 50 nM of siRNA targeting AMPKα1 or AMPKα2. Expression of AMPKα1, AMPKα2, β-catenin, Snail, ZO-1, E-cadherin, and Zeb1 was analyzed by western blot 72 h after transfection. (B) MDA-MB-231 GFP cells were transfected 50 nM siRNA targeting AMPKα1/2 and 100 nM for NTC. Cells were collected 48 h after transfection and injected intravenously into NSG mice. MDA-MB-231 lung metastasis was visualized with fluorescence imaging 36 d later. (C) MDA-MB-231 cells were transfected with siRNA targeting Akt1, AMPKα1, or Akt1/AMPKα1. Expression of Akt1, AMPKα1, cleaved PARP, and Bim was detected 72 h after transfection. (D) MDA-MB-231 GFP cells were transfected with siRNA targeting Akt1, AMPKα1, or Akt1/AMPKα1. Transfection concentrations were: (1) individual siRNA: 50 nM, (2) combination siRNA: 50 nM each (100 nM total), and (3) NTC: 100 nM. Cells were collected 48 h after transfection and injected intravenously into NSG mice. MDA-MB-231 lung metastasis was visualized with fluorescence imaging 40 d later. NTC, Non-targeting control was used as a negative control.