Xenbase Image ID: 137089

Figure 4. USP15 interacts and co-localizes with SMAD6 and ALK3. (a) HEK293 cells were transfected with GFP-USP15 with control vector or mammalian expression vectors encoding N-terminal FLAG-tagged ALK5, ALK3, ALK2 or ALK6. Cells were lysed and extracts (1 mg) subjected to GFP-IPs. GFP-IPs (40%) or extract inputs were resolved by SDS-PAGE and subjected to immunoblotting with the indicated antibodies. (b) HEK293 cells expressing GFP control or GFP-USP15 were transfected with FLAG-ALK3, HA-SMAD6 or both as indicated. Cells were lysed and extracts (1 mg) subjected to GFP-IPs. GFP-IPs (40%) or extract inputs were resolved by SDS-PAGE and subjected to immunoblotting with the indicated antibodies. (c) Fixed cell immunofluorescence was performed on U2OS cells transfected with FLAG-ALK3, HA-SMAD6 and GFP-USP15. Individual and merged pictures are shown, indicating localization of FLAG-ALK3 mainly in the cytosol, HA-SMAD6 in the nucleus and GFP-USP15 in both compartments. GFP-USP15 mainly co-localizes with FLAG-ALK3. Pictures were taken using a 60× lens, scale bar represents 30 μm. Image published in: Herhaus L et al. (2014) . This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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