Additional file 1 of ASC specks exacerbate α‑synuclein pathology via amplifying NLRP3 inflammasome activities
Creators: name: Zheng, Ran
name: Yan, Yiqun
name: Dai, Shaobing
name: Ruan, Yang
name: Chen, Ying
name: Hu, Chenjun
name: Lin, Zhihao
name: Xue, Naijia
name: Song, Zhe
name: Liu, Yi
name: Zhang, Baorong
name: Pu, Jiali
Publisher: name: figshare
Year: 2024
Subjects: subject: Biophysics
subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Molecular Biology
subject: Neuroscience
subject: Immunology
subject: FOS: Clinical medicine
subject: Cancer
subject: Mental Health

description: Additional file 1: Fig. S1 Preparation of sonicated α-synuclein PFFs for stereotaxic injections. Related to Fig. 1. a, b Representative TEM images of recombinant human A53T mutant α-synuclein PFFs before and after sonication. Scale bar: 500 nm. Fig. S2 Propagation of α-synuclein pathology in the mice brains as PD progresses. Related to Fig. 1. a–c Representative immunofluorescence images of P-α-syn (green) in several brain slices of control mice and PFFs-induced PD mice after six- and twelve- weeks treatment. The colored boxes in images are magnified on the right. Fig. S3 No significant protective effects of anti-ASC antibody were found in PFFs-treated PD mice. Related to Fig. 7. a Time schedules of the in vivo experiment in this study. The number of mice in each group: control group, n = 9; PFF group, n = 9; PFF + anti-IgG group, n = 12; PFF + anti-ASC group, n = 10. b Representative images of the tracks of indicated mice in open field test. c, d The total distance traveled and distance traveled in middle zone were recorded and analyzed for each mouse. e The latency to falling during the rotarod test was recorded and analyzed for each mouse. f Representative immunofluorescence staining of TH (green) neurons in the striatum and SNc of mice brains (n = 3–6). White dotted circles indicate the SNc regions. g IB analysis and quantification of TH in bilateral striatum and SN of indicated mice (n = 3). h Representative immunofluorescence staining of Iba1 (green) in the ipsilateral striatum, SN and motor cortex of mice brains (n = 3–6). White dotted squares show the morphology of microglia after magnification. i IB analysis of Iba1 in bilateral striatum and SN of indicated mice (n = 3). Values are presented as mean ± SEM and are analyzed by one-way or two-way ANOVA followed by Tukey’s post hoc test for multiple comparisons. Levels of significance are: *p &lt; 0.05; ns, no significant. Scale bars are as indicated. L, left; R, right; SNc, substantia nigra compacta; SN, substantia nigra. Fig. S4 No differences in α-synuclein pathology and NLRP3 inflammasome activation were observed in brain tissues of anti-ASC antibody treated or anti-isotype treated PD mice. Related to Fig. 7. a Representative immunofluorescence staining of p-α-syn (green) in ipsilateral striatum, SN and cortex of indicated mice (n = 3–6). b–d IB analysis and quantification of SDS-soluble p-α-syn in bilateral striatum and SN tissues (n = 3). e, f IB analysis and quantification of NLRP3 inflammasome signals in bilateral striatum and SN of indicated mice (n = 3). Values are presented as mean ± SEM and are analyzed by two-way ANOVA followed by Tukey’s post hoc test for multiple comparisons. Levels of significance are indicated as follows: *p &lt; 0.05; ns, no significant. Scale bar: 50 μm. L left, R right, SN substantia nigra.