English: In BiFC, WT Phafin2 displayed granular patterns in the cytosol and perinuclear accumulation of the Akt-Phafin2 complex. S1: After Rapamycin treatment to induce autophagy, wild type Phafin2 displayed granular pattern in the cytosol which presumably formation of the Akt-Phafin2 complex at the lysosome and perinuclear accumulation of the Akt-Phafin2 complex determined by the presence of the bright enlarged protein aggregate in BiFC analysis. Method: HeLa cells were transfected by PEI with total of 3 µg of Akt C-term Venus, and N-term Venus Phafin2 along with wild type (WT, movie S1) Phafin2 or PI(3)P interaction defective mutant Phafin2 (R53C, R171A, and R172A, movie S2) as indicated (see details in BiFC experiment of the Material and Methods in supplemental information). The cells were plated on a 12-mm-diameter glass-base dish (Iwaki, 3911-035) in DMEM/F12 (Invitrogen, 110390-021) supplemented with 10% FBS. Twenty four hours after transfection, cells were treated with 10 µM Rapamycin (Sigma) in DMEM/F12 supplemented with 10% FBS for 4 hours in a chamber box on a microscope at 37°C and analyzed using MetaMorph (Version 7.5.6, MDS Analytical Technologies).