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. 2023 Apr 19;9(16):eade1282. doi: 10.1126/sciadv.ade1282

Fig. 5. Astrocytic TDP-43 has cell-autonomous effects on antiviral gene expression, neuroimmune pathways, and chemokine production.

Fig. 5.

(A and B) Western blots of human TDP-43 or mouse and human TDP-43 levels (total TDP-43) in primary hippocampal astrocytes from NTG or ΔNLS mice. TDP-43 levels were normalized to γ-tubulin. Student’s t test with Welch’s correction: ***P < 0.001 versus NTG. (C and D) RNA levels in primary hippocampal astrocytes. Student’s t test: *P < 0.05 and **P < 0.01 versus NTG. (E and F) CXCL10 protein levels in lysates (E) and medium (F) from primary hippocampal astrocytes as measured by enzyme-linked immunosorbent assay (ELISA). **P < 0.01 and *P < 0.05. (G) IFN-γ protein levels in lysates from primary hippocampal astrocytes. *P < 0.05. (H and I) Western blots of phosphorylated and total NF-κB and β-actin in primary hippocampal astrocytes. Student’s t test with Welch’s correction: **P < 0.01. (J and K) Cxcl10 (J) and Cxcl9 (K) mRNA levels in primary hippocampal astrocytes. Vehicle [Veh; dimethyl sulfoxide (DMSO)], Stattic (5 μM, 24 hours), or SN50 (10 μM, 24 hours). One-way ANOVA: F(3,30) = 12.74, P < 0.0001 (Cxcl10) and F(3,28) = 4.39, P = 0.012 (Cxcl9). Dunnett’s post hoc test: ***P < 0.001 and *P < 0.05 versus ΔNLS/vehicle. **P < 0.01. (L and M) Western blot quantification of human TDP-43 (L) and total TDP-43 (M) protein levels in hippocampal astrocytes from ΔNLS mice. Cultures were maintained in DOX (2 μg/ml)–containing or control medium. Student’s t test with Welch’s correction: ***P < 0.001. (N) Cxcl10 mRNA levels in primary hippocampal astrocytes. Some astrocytes were maintained in DOX-containing medium and analyzed at DIV (days in vitro) 8 or 12. Two-way ANOVA: F(1,11) = 14.62, P = 0.0028 for interaction/DIV 8 and F(1,24) = 8.28, P = 0.0083 for interaction/DIV 12. Bonferroni post hoc test: ***P < 0.001. (O) Western blot quantification of phosphorylated NF-κB and STAT3 levels in primary hippocampal astrocytes. Normalized to total levels of each protein. One-way ANOVA: F(2,10) = 29.53, P < 0.0001 (pNF-κB/ΔNLS); F(2,10) = 13.68, P = 0.0014 (pSTAT3/ΔNLS); F(2,6) = 1.77, P = 0.25 (pNF-κB/NTG); and F(2,6) = 0.20, P = 0.83 (pSTAT3/NTG). Dunnett’s post hoc test: ***P < 0.001.