Fig. 5: TFG regulates ULK1 subcellular localization. | Cell Death & Disease

Fig. 5: TFG regulates ULK1 subcellular localization.

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

Fig. 5

A HEK293 cells were transduced with a nontargeting control shRNA (shNC) or TFG-specific shRNAs. Cells were transfected with Flag-ULK1 and HA-TRAF3. Colocalization of overexpressed Flag-ULK1 (green) and HA-TRAF3 (red) were analyzed by confocal microscopy. Nuclear was visualized by DAPI staining (blue). Right panel shows relative fluorescence density of nuclear/cytosol ratio of ULK1 and TRAF3. Data are shown as mean ± s.d. *p < 0.05; n = 3 for each group. B THP-1 cells were transduced with shNC or TFG-specific shRNAs. Nuclear and cytosol fractions from THP-1 cells were extracted and subjected to IB using anti-ULK1 and anti-TRAF3 antibodies. Lower panel shows nuclear/cytosol ratio of ULK1 and TRAF3. Data are shown as mean ± s.d. *p < 0.05; n.s. no significance. C Co-IP assay of endogenous ULK1 and TRAF3 in control or TFG-specific knockdown THP-1 cells. Lower panel shows TRAF3/ULK1 ratio in which indicates binding activity. Data are shown as mean ± s.d. *p < 0.05; n = 3 for each group. Experiments were independently repeated three times.

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