Extended Data Fig. 3: STING signalling drives Sandhoff disease via neuronal death.
From: Innate immune sensing of lysosomal dysfunction drives multiple lysosomal storage disorders
(a, b) Immunofluorescence imaging detected STING aggregation in primary neurons upon 2-hour treatment of STING agonist DMXAA (a) and their quantification (b). Scale bars, 50 μm (left), 10 μm (right). n = 20 cells per group. (c) Immunoblotting revealed that cGAS-STING signalling was activated in primary mixed glial cells upon HSV-I infection. (d, e) Both immunoblotting (d) and qRT-PCR (e) showed that cGAS-STING signalling was activated in primary astrocytes upon DMXAA treatment (n = 3 independent samples). (f, g) TUNEL staining indicated neuronal death (FITC-positive) upon DMXAA treatment for 12-hour and 24-hour (f) and their quantification (g) Scale bars, 50 μm. n = 8 images per group. (h, i) Levels of c-PARP protein showed that prolonged DMXAA treatment was more likely to induce death of primary neurons (h) than glial cells (i). (j) A schematic model of the generation of transgenic mice carrying caSTING in excitatory neurons was shown. (k) Conditional induction of caSTING (STING R281Q) expression in excitatory neurons profoundly activated STING signalling as indicated by IRF3 phosphorylation and accompanied by neuronal loss. (l, m) Nissl staining of mice brains revealed that expression of caSTING in cre-positive mice resulted in neuronal death in the CA1 regions of the hippocampus (l), and their quantification of the neuron density was shown (m). Scale bars, 20 μm. n = 4 mice per group. Error bars in graphs b-m represent mean ± SEM, and P values were calculated by one-way ANOVA with Bonferroni correction. Results in a–m represent three independent experiments.
