Fig 1: Upregulation of PGRN expression in prion-infected mouse brains. (a) qRT-PCR for Grn mRNA from terminally sick RML6-infected C57BL/6J mice (B6 + RML) and age-matched NBH-inoculated C57BL/6J mice (B6 + NBH) and RML6-inoculated Prnp-/- mice (Prnp-/-+RML) (n = 4). Relative expression was normalized to Gapdh expression and represented as percentage of average values in NBH-treated mice. (b) Left, Western blot for PGRN and Actin on brains collected from terminally sick RML6-infected C57BL/6J mice and age-matched NBH-inoculated C57BL/6J mice (n = 4 ~ 5). Right, densitometric quantification of the PGRN Western blot (n = 4 ~ 5). (c) RNA-Seq data of Grn expression in hippocampi at different time points after prion inoculation (n = 3). (d) Ribosomal profiling of Grn expression in CX3CR1-positive microglia at different time points after prion inoculation (n = 3, except n = 2 at terminal stage). (e) RNA-Seq data of Grn expression in sorted microglia (CD11b+) at different time points after prion inoculation (n = 5). (f) Immunofluorescence staining of PGRN on brain sections from RML6- and NBH-inoculated mice. Scale bar: 20 μm. ns: P > 0.05; *: P < 0.05; ***: P < 0.001; ****: P < 0.0001
Fig 2: Microglia specific Grn depletion did not alter prion pathogenesis. (a) qRT-PCR for Grn mRNA and (b) ELISA for PGRN levels in tamoxifen-treated CX3CR1-CreERT;Grnfl/fl and Grnfl/fl mouse brains. n = 5 ~ 6. (c) Survival curves of RML6-infected CX3CR1-CreERT;Grnfl/fl and Grnfl/fl mice untreated or treated with Tamoxifen. The median survival of Grnfl/fl+Tx mice was 172 dpi (n = 25), CX3CR1-CreERT;Grnfl/fl+Tx mice was 170.5 dpi (n = 25), Grnfl/fl-Tx mice was 166 dpi (n = 24), CX3CR1-CreERT;Grnfl/fl-Tx mice was 174dpi (n = 24). There was no significant difference between the groups. (d-g) Left, representative histology of cortex (d), hippocampus (e), thalamus (f) and cerebellum (g) of RML6-infected CX3CR1-CreERT;Grnfl/fl and Grnfl/fl mice treated with Tamoxifen. Scale bar: 100 μm in (d) and (e), 25 μm in (f) and (g). Right, quantification of vacuoles, SAF84 signals, GFAP+ cells and Iba+ cells at various brain regions. n = 6. ns: P > 0.05; *: P < 0.05
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