Journal: bioRxiv

Article Title: Amyloid beta glycation leads to 1 neuronal mitochondrial dysfunction and Alzheimer’s pathogenesis through VDAC1-dependent mtDNA efflux

doi: 10.1101/2024.05.14.594173

Figure Lengend Snippet: ( a ) Immunoblotting analysis of the expression of the cGAS, p-STING, STING, p-TBK1, and p-IRF3, in six-week vehicle or Azeliragon treated APP mice (8-month-old); n = 5. ( b ), Quantification of the expressions of cGAS, p-STING, STING, p-TBK1, and p-IRF3 relative to β-actin. Mean ± SD; n = 5; #P < 0.05, *P < 0.001, one-way ANOVA with Bonferroni’s post hoc test. ( c ), Immunoblotting analysis of the expression of pre-synaptic marker protein synaptophysin and postsynaptic density protein 95 (PSD95) in cortical tissues (including hippocampus) in n = 5. ( d ), Quantification of protein levels of Synaptophysin or PSD95 relative to β-actin. Mean ± SD; n = 5; *P < 0.001, one-way ANOVA with Bonferroni’s post hoc test. ( e ), PLA with anti-dsDNA and anti-VDAC1 antibodies showing the association of VDAC1 protein with cytosolic or nuclear dsDNA in hippocampal CA1 region of the indicated mice. Scale bar, 10 μm. ( f ) Quantification of PLA red dots per cell. Mean ± SD; n = 5, five sections of each brain sample were stained and counted; *P < 0.001, one-way analysis of variance (ANOVA) with Bonferroni’s post hoc test. ( g ), Immunostaining of cGAS with NeuN in Hippocampus of indicated mice. Scale bar, 10 μm. Representative images as shown were from the results of 5 mice in each group. ( h ), Nest score based on nest building behavior. Nesting deficits indicated by incomplete nests and low nest score. Nesting ability indicated by complete nests, high nest score. Mean ± SD; n = 7; *P < 0.01, one-way ANOVA with Bonferroni’s post hoc test. ( i-l ), Escape latency during MWM hidden platform task in indicated mice ( i ), mean number of crossings of the target during the probe test ( j ), time spent in the quadrant with the hidden platform ( k ), and average swim speed (l) of mice in MWM Mean ± SD; n = 7-10; nsP > 0.05, #P < 0.05, *P < 0.01, one-way ANOVA with Bonferroni’s post hoc test.

Article Snippet: The 6.5-month-old APP mice were intraperitoneally (i.p.) injected with Bcl-2-associated X protein (BAX), voltage-dependent anion channel (VDAC), Cyclophilin D (CypD) or RAGE inhibitor i.e., BIP-V5 (5 mg kg−1 BW; 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline; HY-P0081, MCE), NSC 15364 (2.5 mg kg−1 BW; 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline; Cat #HY-108937, MCE), Cyclosporin A (CsA) (1 mg kg−1 BW i.p. 5% DMSO 40% PEG300 5% Tween-80 50% saline; Cat #HY-B0579, MCE) twice each week, and Azeliragon (100 mcg; 10% DMSO 40% PEG300 5% Tween-80 45% Saline; Cat #HY-50682, MCE) every day for 6 weeks.

Techniques: Western Blot, Expressing, Marker, Staining, Immunostaining

Journal: bioRxiv

Article Title: Small molecules inhibit SARS-COV-2 induced aberrant inflammation and viral replication in mice by targeting S100A8/A9-TLR4 axis

doi: 10.1101/2020.09.09.288704

Figure Lengend Snippet: (A) Flow cytometry detecting coronavirus specific neutrophils in the lung and blood of Ifnar -/- mice infected with MHV among Vehicle treated group, Paquinimod treated group, Resatorvid treated group, and Azeliragon treated group at 5 dpi. (B) Flow cytometry detecting coronavirus specific neutrophils in bone marrow and liver of Ifnar -/- mice infected with MHV between control group and Resatorvid treated group at 7 dpi. Mock group means Ifnar -/- mice treated intraperitoneal with Vehicle. (C) qRT-PCR analysis for viral load in the lungs of mice infected intranasally with MHV at 5 dpi between Vehicle treated group and Resatorvid treated group. n = 3 . (D) Flow cytometry detecting coronavirus specific neutrophils in the lungs of hACE2 mice infected with SARS-CoV-2 between control group, Paquinimod and Resatorvid treated group at 5 dpi. (E) qRT-PCR analysis for the expression level of Cxcl2 and S100a8 in the Raw 264.7 cells treated with indicated serum among WT control group, WT Resatorvid treated group, WT Azeliragon treated group and Myd88 -/- group. n = 3. (F)&(G) qRT-PCR analysis for the expression level of several related genes of neutrophil groups in bone marrow of Ifnar -/- mice infected with MHV between control group and Resatorvid treated group at 5 dpi. n = 3. (H) RNA-seq analysis of the level of several related genes of neutrophil groups from the lungs of C57BL/6 mice infected with IAV and hACE2 mice infected with SARS-CoV-2 at 5 dpi. ( *P < 0 . 05; **P < 0 . 01; ***P < 0 . 001; ****P < 0 . 0001 ).

Article Snippet: For the Azeliragon rescue assay, the mice were given intraperitoneally 100 μg/day of Azeliragon (TargetMol; Catalog No. T2507) starting on 2 dpi.

Techniques: Flow Cytometry, Infection, Quantitative RT-PCR, Expressing, RNA Sequencing Assay