Figure 5. PAWS1 interacts with CK1 at the endogenous level GFP pull downs from U2OS PAWS1−/− and PAWS1GFP/GFP cells were resolved by SDS–PAGE, and the gel was stained with Coomassie. Each lane was cut into six pieces, which were subsequently processed for protein identification by mass spectrometry. GFP pull downs from U2OS PAWS1−/− and PAWS1GFP/GFP cells were resolved by SDS–PAGE and analysed by Western blotting using the indicated antibodies. Anti‐CK1α IPs from U2OS PAWS1WT and PAWS1−/− cells were resolved by SDS–PAGE and analysed by Western blotting using the indicated antibodies. U2OS cells were transiently transfected with cDNA encoding FLAG‐tagged PAWS1WT, PAWS1F296A, PAWS1F300A, PAWS1F296A/F300A, PAWS1D262A and FLAG‐empty vector. Anti‐FLAG IPs were immunoblotted with the indicated antibodies. Anti‐CK1α IPs and IgG‐IPs from U2OS PAWS1WT, GFP, PAWS1F296A and PAWS1D262A rescue cells were resolved by SDS–PAGE and analysed by Western blotting using the indicated antibodies. PAWS1 co‐localises with CK1α in U2OS cells. PAWS1−/− cells in which PAWS1WT or PAWS1F296A expression was restored were fixed for immunofluorescence using antibodies against PAWS1 and CK1α. Scale bar is 20 μm. Images from one field of view representative of three biological replicates are included.
Image published in: Bozatzi P et al. (2018)
© 2018 The Authors. 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
Permanent Image Page
Printer Friendly View
XB-IMG-172083