Figure 3. snRNA-Seq analyses in the thalamus and frontal cortex of individuals with and without FTLD-GRN, and Grn^+/+ and Grn^–/– mice.

(A) A schematic diagram for single-cell transcriptomics using the thalamus and frontal cortex from control and FTLD-GRN cases and 19-month-old Grn^+/+ and Grn^–/– mice. (B and C) tSNE plots highlighting the distribution of cell clusters from hTH, hFCX from control and FTLD-GRN cases (B), mTH and mFCX from 19-month-old Grn^+/+ and Grn^–/– mice (C). (D and E) Violin plots showing GRN RNA levels in all the cell types in hTH and hFCX from individuals with and without FTLD-GRN. Statistics uses multiple unpaired Student’s 2-tailed t tests. (F) Immunostains for PGRN in hTH and hFCX of patients with FTLD-GRN. n, neuron; b, blood vessel; g, glia cell; L3, layer 3; L4, layer 4. Scale bars: 100 μm; 20 μm (inset). (G) Quttantification of PGRN intensity in neurons and glia in hTH and hFCX in individuals with and without FTLD-GRN. Statistics uses 2-tailed Student’s t test, n represents the number of independent samples used. (H) Confocal images of PGRN in IBA1⁺ microglia, CD34⁺ endothelial cells, MAP2⁺ neurons, and GFAP⁺ astrocytes, in the hFCX of control and FTLD-GRN patients. Scale bars: 20 μm. (I and J) Gene burden score for each cell cluster in the hTH, hFCX, mTH, and mFCX. Statistics use parametric (mTH, mFCX) or nonparametric (hTH, hFCX) 1-way ANOVA. (K) Bar graphs showing the ratio of the number of nuclei captured in the hTH and hFCX in control and FTLD-GRN patients. Statistics use nonparametric 2-tailed Student’s t test (hTH: ExNeu-1, ExNeu-2, and MG; hFCX: AST-REAC) or parametric 2-tailed Student’s t test (for remaining clusters). ExNeu, excitatiry neurons; InNeu, inhibitory neurons; AST, astrocytes; MG, microglia; OL, oligodendroglia; OPC, oligodendroglial precursors; ENDO, endothelial cells; PER, pericytes. All data represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. NS, not significant.