Fig. 1

DISC1 forms a complex with SNPH. a Co-immunoprecipitation of DISC1 and SNPH. Lysates from CAD cells transfected with EGFP-mDISC1 and mSNPH-Myc constructs were immunoprecipitated with anti-Myc (upper) or anti-GFP (lower) antibodies. Immunoprecipitates were analyzed by western blotting with anti-GFP and anti-Myc antibodies. IP; immunoprecipitation, WB; western blotting. b Co-immunoprecipitation of SNPH deletion mutants and FLAG-mDISC1. A schematic diagram of SNPH domains and SNPH deletion mutants is shown. MBD; microtubule binding domain, ATD; axon targeting domain, MTD; mitochondrial transmembrane domain, TM; transmembrane segment. c Co-immunoprecipitation of FLAG-mDISC1 and SNPH deletion mutants in the mitochondrial transmembrane domain. A schematic diagram of SNPH deletion mutants is also shown. d Co-immunoprecipitation of mSNPH-Myc with EGFP-mDISC1 from the mitochondrial fraction. The mitochondrial and cytosolic fractions from CAD cells transfected with mSNPH-Myc and EGFP-mDISC1 were subjected to immunoprecipitation followed by western blotting. e Co-immunoprecipitation of endogenous SNPH and DISC1 from the mitochondrial fraction of mouse brain tissue. Anti-SNPH immunoprecipitates were analyzed by western blotting with anti-SNPH and anti-DISC1 antibodies. Rabbit IgG was used as negative control for immunoprecipitation. The asterisk and bracket indicate endogenous DISC1 and SNPH, respectively. f Co-localization of EGFP-mDISC1 and mSNPH-Myc in axon of primary cortical neurons at DIV 14 shown by immunocytochemistry. Neurons were transfected as indicated and stained with anti-Myc (cyan) and anti-GFP (green) antibodies. Scale bar; 20 μm