Fig. 2: Development of a transgenic mouse model (NeuLyso-Tag mouse) to isolate neuronal lysosomes from brains.

a A lentiviral vector expressing ^HALAMP1^Myc via the neuron-specific synapsin-1 promoter was used to generate Tg-^HALAMP1^Myc (or “NeuLyso-Tag”) mice via pronuclear injection of linearized DNA comprising the synapsin-1 promoter and the ^HALAMP1^Myc cDNA. b Tg-^HALAMP1^Myc mice were PCR-genotyped using primer-pairs at the N- and the C-terminal ends of the ^HALAMP1^Myc cDNA. Single insertion-site for the transgene was suggested by equal numbers of WT and Tg- ^HALAMP1^Myc pups born per litter (n = 11 litters). There was also no effect of the transgene on birth-ratio and growth (indicated by weight gain; n = 5 per group) of the Tg- ^HALAMP1^Myc mice compared to WT mice. c Immunoblots show that ^HALAMP1^Myc protein is detectable in mouse brains by immunoblots against the N-terminal HA tag and the C-terminal myc tag (representative of n = 7 littermates of each genotype). d Histological colocalization of anti-HA immunofluorescence with NeuN staining in the cortex and hippocampus shows that ^HALAMP1^Myc construct is expressed in the neurons of Tg-^HALAMP1^Myc mouse brains at 1 and 6 months of age (representative of >10 stained sections from n = 3 littermates of each genotype). All data represent means ± SEM. n.s. not significant, by paired 2-tailed Student’s t test for littermate ratio and by RM 2-way ANOVA for weight gain over age (each mouse matched over age). Note: The FSW-^HALAMP1^Myc lentiviral vector is described and tested in Supplementary Fig. S2, and further characterization of the Tg- ^HALAMP1^Myc mice is included in Supplementary Fig. S3.