Fig. 4: Small molecule analogs lead to delayed induction of Grn mRNA and PGRN protein in vitro and in vivo.
A, B Western blot and quantification of Grn+/− mice treated i.p. with analog. A21 (n = 6) and A41 (n = 7) significantly increased PGRN protein in the brain compared to Grn+/− control. A21 and A41 also completely rescued Grn+/− PGRN protein back to Grn+/+ levels while A39 (n = 4) and A40 (n = 4) had no effect. All biological replicates are summarized in mean values ± SD and * represents p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 significance with respect to Grn+/− control (n = 5); # represents p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001 significance with respect to Grn+/+ control (n = 5); ordinary one-way ANOVA with Dunnett’s multiple comparisons test was used to determine significance. C N2A PRGN of A41 treatments at increasing time points (n = 2) with compound and after compound removal. Compound significantly increased PGRN protein and GRN mRNA levels at 18 and 24 h. Statistically significant increased levels of PGRN protein persisted for 12 h after compound removal. Biological replicates are shown as mean values ± SD and ordinary one-way ANOVA with Dunnett’s multiple comparisons was used to determine significance. * represents p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 in respect to DMSO treated control (n = 4). D, E Immunoblot and quantification of Grn+/− mice treated with 50 mg/kg A41 i.p. at increasing time points (n = 3 for each time point). A41 significantly upregulated PGRN protein at 20 and 24 h after treatment compared to controls (n = 6). (24 h data points included in quantification for Fig. 4B) Biological replicates are shown as mean values ± SD and ordinary one-way ANOVA with Dunnett’s multiple comparisons was used to determine significance. * represents p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 in respect to DMSO treated control (n = 7).
