Extended Data Fig. 7: AD4 and oxATP significantly suppressed the phenotypes associated with ARF1 ablation.
Control (Thy1-CreER/Arf1f/+, Arf1+/-) and the neuronal ARF1-KO (Thy1-CreER/Arf1f/f, Arf1-/-) mice were treated with PBS, AD4, and oxATP and assayed for various phenotypes: a, Mitochondrial ROS was directly examined by using MitoSOX from mice with the indicated genotypes and treatments, then stained with MitoSOXTM (red) and MAP2 (green). Scale bars: 5 μm. Data represents one of three independent experiments. b,c, The synapses were checked by staining VGLUT1 and PSD95 (b) and quantified (c) in the spinal cord sections from mice with the indicated genotypes and treatments. (n = 20 images observed from 3 mice). Scale bars: 5 μm. d,e, Immunohistochemical staining (d) and quantification (e) for IBA1 and GFAP in spinal cords from mice with the indicated genotypes and treatments. Scale bar: 50 μm. (n = 8 images observed from 3 mice). f, Immunoblot of GSDMD, N-terminal GSDMD and Caspase11 in spinal cord lysates from control (Arf+/-) and neuronal ARF1-KO (Arf1-/-) mice. GAPDH was used as loading control. Data represents one of three independent experiments. g, The levels of several cytokines, complement C1q and C3 were measured by qRT-PCR in the mouse spinal cord lysates of control and neuronal ARF1-KO mice. (n = 4 mice in each group). h-j, The levels of Tnf and Il1α were measured by qRT-PCR from isolated microglia (h), Astrocytes (i), and neurons (j), in control and neuronal Arf1-KO mice. (n = 4 per group). k-n, The Levels of IL1α (k), IL1β (l), IL18 (m), TNF (n) were measured by ELISA in the mouse spinal cord lysates of control andneuronal ARF1-KO with treatments. (n = 5 per group). Data are represented one of three independent experiments and shown as mean ± SEM. P value was calculated by two-tailed t-test.
