Fig. 5: NXN interrupted the association between DUB3 to bind with Snail and inhibited deubiquitylation of Snail by DUB3. | Cell Death & Disease

Fig. 5: NXN interrupted the association between DUB3 to bind with Snail and inhibited deubiquitylation of Snail by DUB3.

From: NXN suppresses metastasis of hepatocellular carcinoma by promoting degradation of Snail through binding to DUB3

Fig. 5

A Co-immunoprecipitation (co-IP) was performed to detect the interaction between NXN, Snail and DUB3 in 293 T and PLC-8024 cells. NXN did not interact with Snail protein, whereas NXN or Snail could co-immunoprecipitated with DUB3. B DUB3 overexpression reduced the ubiquitinated Snail in 293 T, PLC-8024 and Huh7 cells. C NXN overexpression reduced the DUB3-mediated Snail deubiquitylation in 293T, PLC-8024 and Huh7 cells. Cells were transfected with indicated plasmid. After 48 h, the cells were incubated with MG132 (10 mM) for 4 h, then the cell lysates were immunoprecipitated with anti-Flag, and the immunocomplexes were analyzed by Western blot using an anti-ubiquitin antibody. D NXN overexpression reduced the interaction between DUB3 and Snail. Total lysates of cells with or without NXN overexpression were subjected to IP with anti-DUB3 or anti-Snail Ab, followed by Western blot using the indicated antibodies. The cells were treated with MG132 to inhibit the proteasome.

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