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. Author manuscript; available in PMC: 2020 Oct 7.
Published in final edited form as: Curr Biol. 2019 Sep 5;29(19):3135–3152.e4. doi: 10.1016/j.cub.2019.07.071

Figure 6. Blocking Cell Death in hTDP-43 Expressing Glia Exacerbates Non-Autonomous Neuronal Toxicity.

Figure 6.

(A-F) Distribution of activated Caspase-3 (White) was monitored after pan-glial but post development induction of hTDP-43 at Day 3 (A-C) and Day 4 (D-E) post induction. All glial and neuronal nuclei were co-labeled with antibodies against the Repo (Red) and Elav (Blue) markers. Effects of hTDP-43 on Caspase-3 were monitored when it was co-induced with either nls-LacZ (A, D) control (repots>hTDP-43+nls-LacZ), vs p35 (repots>hTDP-43+p35) or DIAP-1 (repots>hTDP-43+DIAP-1), inhibitors of Caspase-3 function (B,E and C,F, respectively). Scale bar=10μm.

(G) Quantification of total glial numbers from a defined brain region (mean ± SEM). Total glial numbers were 30.5±2.3 (Day 3, n=8) and 8.8±0.9 (Day 4, n=13) in repots>hTDP-43+nls-LacZ, 36.3±1.4 (Day 3, n=16) and 33.5±1.5 (Day 4, n=8) in repots>hTDP-43+p35, and 32.8±1.3 (Day 3, n=15) and 25.1±1.8 (Day 4, n=12) in repots>hTDP-43+DIAP-1. **p<0.01, ****p<0.0001, n.s., no significant difference (Unpaired t-test). ####p<0.0001, n.s., no significant difference (Two-way ANOVA).

(H) Quantification of total neuronal numbers from a defined brain region (mean ± SEM). Total neuronal numbers were 374.4±8.9 (Day 3, n=8) and 311.0±7.2 (Day 4, n=13) in repots>hTDP-43+nls-LacZ, 361.5±9.4 (Day 3, n=16) and 209.3±17.4 (Day 4, n=8) in repots>hTDP-43+p35, and 379.6±6.3 (Day 3, n=15) and 191.6±11.6 (Day 4, n=12) in repots>hTDP-43+DIAP-1. ****p<0.0001 (Unpaired t-test). ####p<0.0001, n.s., no significant difference (Two-way ANOVA).

(I) Quantification of total numbers of Caspase-3 positive glia from a defined brain region (mean ± SEM). Total Caspase-3 positive glial numbers were 17.3±1.0 (Day 3, n=8) and 4.9±0.7 (Day 4, n=13) in repots>hTDP-43+nls-LacZ, 16.3±1.2 (Day 3, n=16) and 22.0±2.1 (Day 4, n=8) in repots>hTDP-43+p35, and 15.9±1.2 (Day 3, n=15) and 11.6±1.1 (Day 4, n=12) in repots>hTDP-43+DIAP-1. *p<0.05, **p<0.01, ****p<0.0001 (Unpaired t-test). ##p<0.01, ####p<0.0001, n.s., no significant difference (Two-way ANOVA).

(J) Quantification of total numbers of Caspase-3 positive neuron (mean ± SEM). Total Caspase-3 positive neuronal numbers were 9.5±1.5 (Day 3, n=8) and 26.5±1.9 (Day 4, n=13) in repots>hTDP-43+nls-LacZ, 18.5±1.3 (Day 3, n=16) and 43.3±3.2 (Day 4, n=8) in repots>hTDP-43+p35, and 19.4±1.6 (Day 3, n=15) and 45.5±2.8 (Day 4, n=12) in repots>hTDP-43+DIAP-1. ****p<0.0001 (Unpaired t-test). #p<0.05, ####p<0.0001 (Two-way ANOVA).

See also Figure S6 and S7.