Fig. 6

A retinitis pigmentosa-causing mutation in the human PDE6G gene causes a frameshift that adds a proline-rich, destabilizing C-terminus with altered protein-protein interactions. a Schematic representation of the wildtype (WT) and mutated (MUT) PDE6G gene. Alternative ss (blue) usage causes a frameshift and translation into the 3′UTR. Strategy for CRISPR/Cas9-mediated insertion of the mutation is shown (see Experimental procedures for details). b Stability of GFP-tagged fl and fs PDE6G C-termini was analyzed as in Fig. 3c. n = 3. c RT-PCR and immunoblotting of either WT Hek293T cells or cells heterozygous for the PDE6G mutation shown in a. d PDE6G +/− cells were treated as indicated and stability of PDE6G isoforms was determined as in Fig. 3c using a PDE6G specific antibody, n = 3. e Specific co-precipitation of PDE6α/β-Flag with GFP-PDE6G-fl but not GFP-PDE6G-fs. The N-terminus of the unrelated RNA helicase Brr2 served as Flag-tagged control protein. * marks IgG