Figure 1
From: Neuralized family member NEURL1 is a ubiquitin ligase for the cGMP-specific phosphodiesterase 9A
NEURL1 and NEURL1B interact with PDE9A. (A) PDE9A-V5His was expressed alone or together with either NEURL1-Flag, Flag-NEURL1 or Flag-NEURL1B in HEK293-FT cells. (B) PDE9A-V5His was expressed alone or together with untagged NEURL1 in HEK293-FT cells. In A and B the proteins were immunoprecipitated and immunoblotted using the indicated antibodies (untagged NEURL1 was detected with a custom-raised polyclonal NEURL1 antibody). (C) PDE9A-V5His and Flag-NEURL1 or NEURL-Flag or Flag-NEURL1B were co-expressed in HEK293 cells and their localisation was visualised by indirect immunofluorescence using respective tag-specific antibodies (co-localisation of green and red signals results in yellow additive colour in the Merge column). Nuclei were visualised using Hoechst 33342 (blue). (D) Flag-NEURL1 and E2-tagged PDE5A, PDE9A and PDE11A were expressed alone or in combination in HEK293-FT cells. Cell lysates were immunoprecipitated and immunoblotted with the indicated antibodies. IB, immunoblot; IP, immunoprecipitation.
