Journal: bioRxiv

Article Title: Cryo-EM structures of α-synuclein filaments from Parkinson’s disease and dementia with Lewy bodies

doi: 10.1101/2022.07.12.499706

Figure Lengend Snippet: Sections from brain regions contralateral to those used for cryo-EM structure determination were stained with monoclonal antibody Syn1 (1:1,000). (a), Cingulate cortex from PD; (b), Cingulate cortex from PDD1; (c), Cingulate cortex from PDD2; (d), Frontal cortex from DLB1; (e), Frontal cortex from DLB2; (f), Cingulate cortex from DLB3. Scale bars: a-c, f, 100 μm; d,e, 50 μm.

Article Snippet: Before blocking, membranes were fixed with 1% paraformaldehyde for 30 min. Primary antibodies were: Syn303 [a mouse monoclonal antibody that recognizes residues 1-5 of human α-synuclein ( )] (BioLegend) at 1:4,000, Syn1 [a mouse monoclonal antibody that recognizes residues 91-99 from the NAC region of human α-synuclein ( )] (BD Biosciences) at 1:4,000 and PER4 at 1:4,000.

Techniques: Cryo-EM Sample Prep, Staining

Journal: bioRxiv

Article Title: Cryo-EM structures of α-synuclein filaments from Parkinson’s disease and dementia with Lewy bodies

doi: 10.1101/2022.07.12.499706

Figure Lengend Snippet: PER4 was used at 1:50 in (a-c). (a), PD (Cingulate cortex); (b), PDD1 (Cingulate cortex); (c), DLB3 (Cingulate cortex); Syn303, Syn1 and PER4 were used at 1:4,000 in (d-f). The brain regions used for cryo-EM were also used for immunoblotting. The arrow points to the position of monomeric α-synuclein.

Article Snippet: Before blocking, membranes were fixed with 1% paraformaldehyde for 30 min. Primary antibodies were: Syn303 [a mouse monoclonal antibody that recognizes residues 1-5 of human α-synuclein ( )] (BioLegend) at 1:4,000, Syn1 [a mouse monoclonal antibody that recognizes residues 91-99 from the NAC region of human α-synuclein ( )] (BD Biosciences) at 1:4,000 and PER4 at 1:4,000.

Techniques: Cryo-EM Sample Prep, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: High-Salt Diet Accelerates Neuron Loss and Anxiety in APP/PS1 Mice Through Serpina3n

doi: 10.3390/ijms252111731

Figure Lengend Snippet: HS diet induces neuron reduction in AD mice. ( A ) Thioflavin T staining was used to detect S-plaque in mice brains. No S-plaque was observed in the HS/ND-treated wt mice; n = 5 in each group. Scale bar = 50 μm. ( B , C ) Nissl staining was performed to detect the neuron number of HS/ND-treated wt mice. Neuron numbers were counted in the CA1 region of the hippocampus in each group. Scale bar = 50 μm. ( D , E ) Thioflavin T staining was used to detect S-plaque in the brains of HS/ND-treated AD mice. The number of S-plaques in parietal cortex, hippocampus, and prefrontal cortex was counted; n = 5 in each group. Scale bar = 50 μm. ( F ) Representative Nissl staining of hippocampus in each group. CA1 regions are magnified on the right. The immunohistochemistry for NeuN in the hippocampus of each group is also shown. Between the two green dotted lines is the neurons in the CA1 region. Scale bar = 50 μm. ( G ) The Nissl body-positive neuron numbers were counted from the CA1 region of brain slides from five mice in each group; n = 5 in each group. The scatter bar chart shows the integrated density of NeuN-positive particles in each brain slide from five mice, which was measured using Image J software; n = 5 in each group. ( H ) Immunohistochemistry for syn1 staining in the hippocampus of each group. Between the two green dotted lines is the neurons in the Or, Rad, and Mol regions; scale bar = 20 μm. ( I ) The scatter bar chart shows the integrated density of Syn1-positive particles from Or, Rad, and Mol, respectively. Each brain slide from five mice was measured using Image J software; n = 5 in each group. ( J , K ) Western blot analysis for caspase 3, Bax, Syn1, and β-tubulin 3; β-actin was used as an internal control. The data are the mean ± SE; n = 4 biologically independent animals. Blue represent the ND treated mice, red represent the HS treated mice.

Article Snippet: The sections were incubated with rabbit anti-NeuN (1:500, 14H6L24, Invitrogen, CA, USA) or mouse anti-syn1 (1:200, 7H10G6, Novus, CA, USA) overnight at 4 °C.

Techniques: Staining, Immunohistochemistry, Software, Western Blot, Control

Journal: Cell reports. Medicine

Article Title: Increased intestinal bile acid absorption contributes to age-related cognitive impairment.

doi: 10.1016/j.xcrm.2024.101543

Figure Lengend Snippet: Figure 2. The excess of CPBA and ammonia in the brain is associated with hippocampal synapse loss in aging rodents (A‒D and I‒L) The percentages of CPBAs and ammonia level as well as the CPBA composition (e.g., taurocholic acid [TCA]) in the brain of rodents in both genders (n = 6–8/group). (E, F, M, and N) The representative images and counts of hippocampal synapses in rats and mice resulted from transmission electron microscopy (n = 6–8/group, 3 repeats/sample). (G and O) The concentrations of hippocampal Synapsin 1 (Syn1) in both genders of rodents (n = 68/group, 3 repeats/sample). (H and P) Pearson’s r correlation between hippo- campal Syn1 level and brain TCA or ammonia in rodents. The comparisons of CPBA, ammonia, and synapse data in rodents were performed with two-way ANOVA. The statistical difference is labeled as follows: ***p < 0.005 vs. 24-week female (F24); ##p < 0.01; ###p < 0.005 vs. 24-week male (M24); $$p < 0.01 vs. 111-week male (M111) for the comparison of the CBPA composition among groups; *p < 0.05; **p < 0.01; ***p < 0.001 for other comparisons. See also Figures S2 and S3.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-SYN1 Proteintech Cat#20258-1-AP, RRID: AB_2800493 Mouse monoclonal anti-PSD95 Abcam Cat#20665-1-AP, RRID: AB_300453 Rabbit monoclonal anti-b-Actin Cell Signaling Technology Cat#4970, RRID: AB_2223172 Anti-rabbit, HRP-linked Cell Signaling Technology Cat#7074, RRID: AB_2099233 Anti-mouse, HRP-linked Cell Signaling Technology Cat#7076, RRID: AB_330924 Anti-rabbit, Alex Flour 488-linked Abcam Cat#ab150077, RRID: AB_2630356 Anti-mouse, Alex Flour 647-linked Abcam Cat#ab150179, RRID: AB_2884038 Biological samples Serum from mice This paper N/A Tissues from mice This paper N/A Chemicals, peptides, and recombinant proteins Phosphate buffered saline Sigma Cat#P2272, CAS: NA Paraformaldehyde Sigma Cat#158127, CAS: 30525-89-4 RIPA buffer Beyotime Biotechnology Cat#P0013C, CAS: NA DNase I Sigma Cat#11284932001, CAS: NA Papain Sigma Cat#232-627-2, CAS: 9001-73-4 B-27 Gibco Cat#17504044 L-glutamine Gibco Cat#25030149 Critical commercial assays Asbt Elisa kit (for rat) Jiangsu Mei Biao Biological Technology Co., Ltd. Cat#MB-7410A Asbt Elisa kit (for mouse) Jiangsu Mei Biao Biological Technology Co., Ltd. Cat#MB-6453A Syn1 Elisa kit (for rat) S.A.B Cat#EK3880 Syn1 Elisa kit (for mouse) S.A.B Cat#EK6728-2 Ammonia Assay Kit Sigma-Aldrich Cat#AA0100, SYBR Green PCR Master Mix Vazyme Cat#A25742 Pierce BCA Protein Assay Kit Thermo Fisher Cat#23227 Clarity Western ECL Substrate Bio-Rad Cat#1705060 Experimental models: Cell lines Rat primary hippocampal neurons This paper N/A Experimental models: Organisms/strains Wistar rat Vital River laboratory animal technology Co. Ltd., Beijing, China N/A C57BL/6J mouse Shanghai SLAC Laboratory Animal Co., Ltd N/A Software and algorithms GraphPad Prism 9.3 software GraphPad Software http://www.graphpad.com/ Adobe Illustrator Adobe https://www.adobe.com/products/illustrator.html TargetLynx 4.2 Waters Corporation N/A Microsoft 365 Microsoft https://www.microsoft.com/zh-cn/ microsoft-365/buy/compare-allmicrosoft-365-products?tab=1 (Continued on next page) e1 Cell Reports Medicine 5, 101543, May 21, 2024

Techniques: Transmission Assay, Electron Microscopy, Labeling, Comparison