Fig. 7

Delaying vision loss in early-onset RP model mice via viral gene delivery of anti-Prox1 antibody.

a Retinal sections from a healthy 83-year-old donor eye and a 79-year-old RP patient eye were stained with rabbit anti-Prox1, goat anti-Sox2, and mouse anti-Rhodopsin antibodies. Nuclei of the cells in the retinal sections were visualized by DAPI staining. Sox2-positive MG nuclei are outlined by dotted-lines. b The thickness of the ONL in the healthy and RP patient retina was compared to the thickness of the INL in the same section, and the relative values are presented in the graph. Numbers of samples analyzed are shown in the graph (data from 6 independent retinal sections from 2 eyes). ****, p < 0.001 (one-sided Student’s t-test) (c) Relative Prox1 immunofluorescent intensity in MG in the corresponding retina, normalized to Prox1 intensity in BCs within the same image, is shown. Each dot represents the median intensity collected from one retina. Numbers of samples analyzed are shown in the graph (data from 12 independent retinal sections from 2 eyes). *** p < 0.005; **** p < 0.001 (one-sided Student’s t-test). d The distribution of Prox1 in the retinas of mice with heterozygous and homozygous rd10 mutations in Pde6b gene (Pde6b^rd10) was investigated by immunostaining. MG cells among Prox1-positive cells were determined by co-staining of Prox1 and Sox2. Boxed areas in top row are magnified in the next three rows. Sox2-positive MG nuclei are outlined by dotted-lines. e Prox1 intensity in Sox2-positive MG relative to other retinal neurons in the same image is presented in the graph. Numbers of samples analyzed are shown in the graph (data from 4 independent litters). ** p < 0.01; *** p < 0.005; **** p < 0.001 (one-sided Student’s t-test). fPde6b^rd10/rd10;Glast-CreERT;R26^tdTom/+ mice were intravitreally injected with AAV2-Ctrl Ab or AAV2-αProx1 at P10, followed by Tam injection to activate the CreERT, resulting in the expression of tdTom Cre reporter in MG. Mice were also injected with EdU daily from P33 to P38 to label cells born during last 6 days before sample collection at P39. g The identities of tdTom-expressing MG cell lineage cells in mouse retinas were investigated by co-immunostaining for cell type-specific markers. Immunostaining images are provided in Supplementary Fig. 15f, except for the rPR marker, Rhodopsin. The birth of these cells between P33 and P39 was determined by EdU-labeling. Boxed areas in top row are magnified in the next rows. White arrowhead points EdU-labeled cell rPR nuclei and black arrowhead indicates EdU-labeled cell nucleus in the choroid. h Mean numbers of cells expressing corresponding markers in the retinal area are provide in the graph. Numbers of samples analyzed are shown in the graph (data collected from 4 independent litters). i Visual acuities of the mice measured at the indicated postnatal days. *, p < 0.05; **, p < 0.01 (one-sided Student’s t-test) Error bars in the graphs represent SEM (n = 8; 5 independent litters).