Fig. 5.

Immunofluorescence analyses of N-PRβ-KO and N-PRβ-KO-MC mice.

(a) In an immunofluorescence analysis, PDGFRα (green) is expressed on GFAP-positive astrocytes (red) in WT and N-PRβ-KO mice at 4 weeks. However, astrocytes of N-PRβ-KO mice are hypertrophic compared with WT mice. Hoechst-stained nuclei (blue). (b) Based on an immunofluorescence analysis, PDGFRβ (green) was expressed on both αSMA⁻ and αSMA⁺ pericytes (red) in WT mice at 4 weeks. Because NG2⁺αSMA⁺ microvascular pericyte recruitment is observed in N-PRβ-KO mice, PDGFRβ (green) is preferentially expressed on αSMA⁺ pericytes (red) at 4 weeks. Arrowheads indicate αSMA⁺ pericytes detached from blood vessels suppressing PDGFRβ expression. (c) Schematic representation of the transgenic and mutated alleles of N-PRβ-KO-MC. Exons 4 to 7 of Pdgfrb flanked by two loxP sites are deleted by nlsCre activity driven by the Nestin promoter. At the same time, mCherry expression is driven by the Rosa26 promoter, when two loxP sites flanking the stop codon are deleted by nlsCre activity. (d) In N-PRβ-KO-MC retinas at 2 weeks, αSMA⁺ pericytes detached from blood vessels (green, indicated by green-arrowheads) never express mCherry. Some mCherry⁺ cells (red, indicated by magenta-arrowheads) are observed around the blood vessels (cyan), but not express αSMA. Hoechst-stained nuclei (blue). Scale bar = 100 μm (a and b), 50 μm (d).