Fig. 6: TFG suppresses TRAF3-mediated ULK1 degradation. | Cell Death & Disease

Fig. 6: TFG suppresses TRAF3-mediated ULK1 degradation.

From: TRK-fused gene (TFG) regulates ULK1 stability via TRAF3-mediated ubiquitination and protects macrophages from LPS-induced pyroptosis

Fig. 6

A Flag-ULK1 and V5-TFG were transfected into HEK293 cells together with HA-Ub-K48 or HA-Ub K48R as indicated. Flag-ULK1 was isolated by IP and subjected to IB assays using anti-HA and anti-Flag antibodies. Protein lysates were also subjected to direct IB (bottom panels). B Flag-ULK1 and HA-Ub K48 were transfected into HEK293 cells together with V5-tagged wild-type TFG or mutant TFG (Δ11–91, lacking TRAF3 binding domain). Flag-ULK1 were isolated by IP and subjected to IB using anti-HA and anti-Flag antibodies. Protein lysates were also subjected to direct IB (bottom panels). C THP-1 cells were transduced by shNC, shTFG, and/or shTRAF3. Total cell lysates were extracted and ULK1 protein level were analyzed by IB assay. D A proposed model of TFG regulates TRAF3-mediated ULK1 ubiquitination and proteasome degradation. Experiments were independently repeated three times.

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