Fig. 3: TFG deficiency facilitates ULK1 ubiquitination and proteasome degradation.

A THP-1 cells were transduced by shNC, shTFG#1, and shTFG#2 and treated with MG132 for 2 h before harvest. ULK1 was isolated by IP and subjected to IB assay using anti-ubiquitin (Ub) and anti-ULK1 antibodies. Whole-cell lysates were also subjected to direct IB (bottom panels). B THP-1 cells transduced with shNC, shTFG, shTRAF3, and shTFG+shTRAF3 were treated with MG132 for 2 h before harvest. ULK1 was isolated by IP and subjected to IB assays using anti-Ub and anti-ULK1 antibodies. C V5-TFG, Flag-ULK1, and HA-Ub were transfected into shNC- or shTRAF3-transduced HEK293 cells as indicated. Flag-ULK1 were isolated by IP and subjected to IB assays as indicated. Protein lysates were also subjected to direct IB (bottom panels). Experiments were independently repeated three times.