Additional file 1 of Liver-specific adiponectin gene therapy suppresses microglial NLRP3-inflammasome activation for treating Alzheimer’s disease
Creators: name: Ng, Roy Chun-Laam
name: Jian, Min
name: Ma, Oscar Ka-Fai
name: Xiang, Ariya Weiman
name: Bunting, Myriam
name: Kwan, Jason Shing-Cheong
name: Wong, Curtis Wai-Kin
name: Yick, Leung-Wah
name: Chung, Sookja Kim
name: Lam, Karen Siu-Ling
name: Alexander, Ian E.
name: Xu, Aimin
name: Chan, Koon-Ho
Publisher: name: figshare
Year: 2024
Subjects: subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Neuroscience
subject: Pharmacology
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Cancer
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Virology

description: Additional file 1: Figure S1. Adiponectin deficiency exacerbates microgliosis in the early adult stage but not in aged AD mice. A Immunofluorescent staining of Iba1 indicates the changes of spatiotemporal microgliosis in the hippocampus of wildtype, APN−/−, 5xFAD, and APN−/−5xFAD mice at different age (3, 6, 9, 12, 15, 18-month) (n = 3 mice for each experimental group) (Magnification 10 × 20; Scale bar: 50μm). (B) Relative intensity of microglial Iba1 staining in the hippocampus of wildtype, APN−/−, 5xFAD, and APN−/−5xFAD mice at different age (3, 6, 9, 12, 15, 18-month). Data were presented as the mean ± s.e.m. n.s. not significant; *p &lt; 0.05; **p &lt; 0.01; ***p &lt; 0.001. Statistical analysis was performed by one-way ANOVA followed with Bonferroni’s post hoc comparison tests. Figure S2. No mononuclear cell infiltration into the liver after AAV administration. H &amp; E staining of the liver in wildtype, AAV2/8-GFP-treated, and AAV2/8-APNTri-treated 5xFAD indicated no infiltration of immune cell in the liver of AAV-treated mice. (n = 4 mice for each experimental group) (Magnification: 10 × 10; Scale bar: 50μm). Figure S3. AAV2/8-APNTri treatment exerts no significant effect in motor functions and increases level of synaptic proteins in the brain of AD mice. A Total distance travelled of wildtype, AAV2/8-GFP-treated and AAV2/8-APNTri-treated 5xFAD in the visible platform of Morris Water Maze. (n = 9 mice for wildtype and n = 11 for both AAV-treated groups). B Average swim speed of wildtype, AAV2/8-GFP-treated and AAV2/8-APNTri-treated 5xFAD in the visible platform of Morris Water Maze. (n = 9 mice for wildtype and n = 11 for both AAV-treated groups). C Western blotting assay of synaptophysin and spinophilin in the hippocampus of AAV2/8-GFP-treated and AAV2/8-APNTri-treated 5xFAD. (n = 5 mice for each experimental group). D Densitometric analysis of synaptophysin and spinophilin in the hippocampus of AAV2/8-GFP-treated and AAV2/8-APNTri-treated 5xFAD. Data were presented as the mean ± s.e.m. n.s. not significant; *p &lt; 0.05; **p &lt; 0.01; ***p &lt; 0.001. Statistical analysis (A, B) were performed by one-way ANOVA followed with Bonferroni’s post hoc comparison tests and (D) was performed by unpaired t test. Figure S4. AAV2/8-APNTri-treated reduces fibrillar amyloid deposition in the cortex and astrogliosis in the cortex and hippocampus of 5xFAD mice. A Thioflavin staining of Aβ (green) in the cortex of AAV2/8-GFP-treated (n = 6), and AAV2/8-APNTri-treated 5xFAD mice (n = 6). (Magnification: 10 × 20; Scale bar: 50μm). B Relative intensity of the thioflavin-stained Aβ cortex of AAV2/8-GFP-treated, and AAV2/8-APNTri-treated 5xFAD mice. C GFAP (red) and thioflavin S (green) double-immunofluorescent staining in the hippocampus and cortex of AAV2/8-GFP-treated (n = 6), and AAV2/8-APNTri-treated 5xFAD mice (n = 6). (Scale bar: 50μm). D Relative intensity of GFAP in the hippocampus and cortex of AAV2/8-GFP-treated, and AAV2/8-APNTri-treated 5xFAD mice. Data were presented as the mean ± s.e.m. n.s. not significant; *p &lt; 0.05; **p &lt; 0.01; ***p &lt; 0.001. Statistical analyses were performed by unpaired t test. Table S1. Recombinant AAV2/8 did not cause significant changes of body weight, weight changes, and liver weight 20 weeks after administration. Wildtype (n = 9), 5xFAD (n = 7), AAV2/8-eGFP treated 5xFAD (n = 11), and AAV2/8-APNTri treated 5xFAD (n = 11).