Journal: bioRxiv

Article Title: The Endothelial Cell-Expressed Prion Protein Prnd/Doppel Promotes Neovascularization and Long-term Recovery after Ischemic Stroke

doi: 10.64898/2026.02.21.707233

Figure Lengend Snippet: (A); Left panels: Representative immunofluorescence images showing TMEM119 (microglia marker), CD68 (macrophage/activated microglia marker) and NeuN (mature neuronal marker) staining in the ipsilateral cortex of Prnd WT and KO mice at 30 days post-MCAO. CD68 expression is elevated in Prnd KO mice, indicating persistent neuroinflammation. NeuN-positive neurons are reduced in Prnd KO mice, demonstrating decreased neuronal survival. Scale bars represent 50 μm. Right panel: Quantification of NeuN-positive cells in the ipsilateral cortex reveals significantly fewer surviving neurons in Prnd KO mice compared to WT controls at 30 days post-MCAO (n = 3 mice per group). Data are presented as means ± SEM; *p < 0.05 using two-tailed Student’s t-test. (B); Quantitative morphometric analysis of cerebrovascular networks in the ipsilateral cortex at 30 days post-MCAO. Parameters assessed include vessel area, vessel percentage area, total vessel length, and total number of junctions. All parameters are significantly increased in Prnd KO mice; however, qualitative assessment reveals severely disrupted vascular architecture compared to the relatively organized network in WT mice, indicating impaired functional vascular recovery (n = 4 mice per group). Data are presented as means ± SEM; *p < 0.05 using two-tailed Student’s t-test.

Article Snippet: We added primary antibodies: Prnd (Thermo Fisher Scientific, PIPA5113502), VEGF (Thermo Fisher Scientific, ENP802), CD31 (R&D Systems, AF3628), ZO-1 (Thermo Fisher Scientific, PIPA585256 ), Claudin 5 (Thermo Fisher Scientific, 35-2500), GFAP (Novus, NBP1-05198), CD68 (Novus, NBP2-33337), Neun (Thermo Fisher Scientific, PIPA5143552), TMEM 119 (Novus Biologicals, NBP230551), Laminin (Sigma, L9393), and VE-Cadherin (BD Pharmingen, 550548) to the slides and incubated the slides overnight at 4°C in a moisturized dark box, washed three times (5min per wash with gentle rocking at room temperature) and added secondary antibodies (1:500 dilutions) and incubated at room temperature for 1 hr.

Techniques: Immunofluorescence, Marker, Staining, Expressing, Two Tailed Test, Functional Assay

Journal: bioRxiv

Article Title: The Endothelial Cell-Expressed Prion Protein Prnd/Doppel Promotes Neovascularization and Long-term Recovery after Ischemic Stroke

doi: 10.64898/2026.02.21.707233

Figure Lengend Snippet: (A); Representative immunofluorescence images showing ms IgG, TMEM119 (resident microglia marker), CD68 (macrophage/activated microglia marker), and NeuN (mature neuronal marker) expression in the ipsilateral cortex of RD control and Pcre-RD mice at 14 days post-MCAO. Enhanced NeuN staining in Pcre-RD mice indicates improved neuronal survival during the subacute recovery phase (n = 3 mice per group). Scale bars represent 100 μm. (B); Quantification of NeuN-positive neurons in the ipsilateral cortex at 14 days post-MCAO. Pcre-RD mice exhibit significantly more surviving neurons compared to RD controls, demonstrating that endothelial Prnd activation confers neuroprotection during the recovery phase (n = 3 mice per group). Data are presented as means ± SEM; *p < 0.05 using two-tailed Student’s t-test. (C); Representative immunofluorescence images showing ms IgG, TMEM119, CD68, and NeuN expression in the ipsilateral cortex of RD and Pcre-RD mice at 30 days post-MCAO, during the chronic recovery phase (n = 3 mice per group). Scale bars represent 50 μm. (D); Quantification of NeuN-positive neurons in the ipsilateral cortex at 30 days post-MCAO. Pcre-RD mice show a trend toward increased neuronal survival compared to RD controls, though the difference does not reach statistical significance at this later time point (n = 3 mice per group). Data are presented as means ± SEM using two-tailed Student’s t-test. These results suggest that the neuroprotective benefits of Prnd activation are most pronounced during the subacute recovery phase.

Article Snippet: We added primary antibodies: Prnd (Thermo Fisher Scientific, PIPA5113502), VEGF (Thermo Fisher Scientific, ENP802), CD31 (R&D Systems, AF3628), ZO-1 (Thermo Fisher Scientific, PIPA585256 ), Claudin 5 (Thermo Fisher Scientific, 35-2500), GFAP (Novus, NBP1-05198), CD68 (Novus, NBP2-33337), Neun (Thermo Fisher Scientific, PIPA5143552), TMEM 119 (Novus Biologicals, NBP230551), Laminin (Sigma, L9393), and VE-Cadherin (BD Pharmingen, 550548) to the slides and incubated the slides overnight at 4°C in a moisturized dark box, washed three times (5min per wash with gentle rocking at room temperature) and added secondary antibodies (1:500 dilutions) and incubated at room temperature for 1 hr.

Techniques: Immunofluorescence, Marker, Expressing, Control, Staining, Activation Assay, Two Tailed Test