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Fig. 5 | Journal of Biomedical Science

Fig. 5

From: NEDD4 regulates ubiquitination and stability of the cell adhesion molecule IGPR-1 via lysosomal pathway

Fig. 5

NEDD4 promotes lysosomal degradation of IGPR-1. a HEK-293 cells expressing IGPR-1 were transfected with an empty vector or wild-type NEDD4-HA or WW domain truncated NEDD4, where all the four WW domains was deleted (ΔWW-NEDD4). Cells were lysed and whole cell lysates were immunoblotted with an anti-IGPR-1 antibody, anti-HA antibody or Anti-Myc antibody for ΔWW-NEDD4. The membrane was also blotted for GAPDH for protein loading control. b HEK-293 cells expressing IGPR-1 were treated with MG132 or bortezomib for two hours, cells were lysed and whole cell lysates were blotted for IGPR-1 or GAPDH. c HEK-293 cells expressing IGPR-1 were treated with control vehicle, bafilomycine or NH4Cl for 18 h. Cells were lysed and whole cell lysates were blotted for IGPR-1 or GAPDH. d HEK-293 cells expressing IGPR-1 were transfected with an empty vector or wild-type NEDD4-HA. After 46 h of transfection, cells were with control vehicle or bafilomycine and incubated for additional 18 h. Cells were lysed and whole cell lysates were blotted for IGPR-1 or GAPDH. e Melanoma cell lines (MEL-JoSo, SK-Mel-28 and A375) were treated with control vehicle or bafilomycine for 18 h. Cells were lysed and whole cell lysates were blotted for IGPR-1 or GAPDH. The graphs ae are representative of three independent experiments. P < 0.05

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