Fig. 1: Protein expression of VPS35 is not altered in SN of heterozygous VPS35^D620N/+ knockin mouse.

A Heterozygous VPS35^D620N/+ mouse was identified by performing PCR assays using tail DNA of wild-type (WT) or VPS35^D620N/+ mouse. Expected sizes of PCR products for WT allele and VPS35^D620N allele were 660 and 710 bp, respectively. B Following RT-PCR reaction using total RNA extracted from SN of WT or VPS35^D620N/+ mouse, DNA sequencing showed that in contrast to normal GAT (D620) sequence of VPS35 in WT mouse, both mutated AAT (N620) and wild-type GAT (D620) sequences of VPS35 were found in heterozygous VPS35^D620N/+ mouse. C Following RT-PCR reaction, a similar level of mRNA, which encodes full-length coding region of VPS35 mRNA, was observed in SN of WT or VPS35^D620N/+ mouse. Real-time quantitative RT-PCR assays showed that VPS35 mRNA level in SN of VPS35^D620N/+ knockin mice was not significantly different from that of WT mice. Each bar shows mean ± S. E. value of four mice. D Protein level of VPS35 in SN of VPS35^D620N/+ knockin mouse was similar to that of VPS35 in SN of WT mouse. Each bar represents mean ± S. E. value of five mice. E WT and VPS35^D620N/+ mice exhibited similar body weight–age curves. Each point shows mean ± S. E. value of 30–35 mice. F WT mice and heterozygous VPS35^D620N/+ mice displayed similar Kaplan–Meier survival curves (WT mice, n = 30; VPS35^D620N/+ mice, n = 30).