Journal: Cells

Article Title: Avenanthramide-C as Alzheimer's Disease-Modifying Therapy: Early and Sustained Intervention Prevents Disease Progression in Mouse Models.

doi: 10.3390/cells14110826

Figure Lengend Snippet: Figure 2. Long-term Avn-C oral administration initiated at the early AD stage lowers Aβ1-42 production and prevents amyloid-mediated disease advancement in AD mouse models. (a) Western blot and quantitative analysis of BACE1, APP, and Aβ1-42 levels from the hippocampal lysate of WT, 5xFAD (vehicle and Avn-C group) mice (n = 4). A severe amyloid-mediated disease progression was observed from the vehicle 5xFAD group with increased APP, BACE1, and Aβ1-42 levels compared to the WT mice, and Avn-C lowers these protein expressions and halts amyloid progression in 5xFAD mice orally administered with Avn-C three months from early AD stage. (b) Fluorescent confocal z-stack image of Aβ1-42 (green), DAPI (blue) from Tg2576 vehicle and 3 months Avn-C treated mice hippocampal sections (2 sections/group) focused on CA3 region (scale bar 20 µm). (c) Quantitative analysis of Aβ42 fluorescent intensities are shown in the graph (n = 3), where Aβ1-42 levels at the CA3 hippocampal region were lowered by the Avn-C administered early at AD stage with three months of continuous treatment. In all panels, the error bar indicates S.E.M. Overall differences among groups were analyzed using the unpaired t-test two-tailed, one-way ANOVA (*** p < 0.001), followed by pairwise comparisons between groups using post hoc Tukey’s test. Statistical significance is indicated as *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: Slides were mounted with ProLong Gold Antifade reagent with DAPI (Cell Signaling, Danvers, MA, USA).

Techniques: Western Blot, Biomarker Discovery, Two Tailed Test

Journal: Cells

Article Title: Avenanthramide-C as Alzheimer's Disease-Modifying Therapy: Early and Sustained Intervention Prevents Disease Progression in Mouse Models.

doi: 10.3390/cells14110826

Figure Lengend Snippet: Figure 5. Early AD-stage Avn-C administration with long-term treatment prevents chronic activation and reduces the cell body (soma) size of microglial cells in AD mouse models. (a–c) The confocal z-stack fluorescent image Iba1 (red) microglial marker and DAPI (blue) nuclei. Quantitively, cell area (soma, yellow arrow) and fluorescent intensity are measured from 5xFAD vehicle and 3 months Avn-C orally administered mice hippocampal sections (2 sections/mouse) (n = 4); for cell soma size from hippocampal sections, 6–7 cells/sections were analyzed (scale bar 20 µm). (d) Western blot and quantitative analysis of IBA1 microglial marker from WT, vehicle, and 3 months Avn-C-treated (5xFAD (n = 3) and Tg2576 (n = 4)) mice hippocampal lysate. Mean ± S.E.M.s. Overall differences among groups were analyzed using unpaired t-test two-tailed, one-way ANOVA (*** p < 0.001), and pairwise comparisons between groups were performed using post hoc Tukey’s test. Statistical significance is indicated as *** p < 0.001, ** p < 0.01, ns p > 0.05 (no significance).

Article Snippet: Slides were mounted with ProLong Gold Antifade reagent with DAPI (Cell Signaling, Danvers, MA, USA).

Techniques: Activation Assay, Marker, Western Blot, Two Tailed Test

Journal: Cells

Article Title: Avenanthramide-C as Alzheimer's Disease-Modifying Therapy: Early and Sustained Intervention Prevents Disease Progression in Mouse Models.

doi: 10.3390/cells14110826

Figure Lengend Snippet: Figure 6. Long-term oral administration of Avn-C from the early AD stage reduces large amyloid plaque deposition, strengthens the microglial cell barrier in hippocampal tissue, and protects and enhances microglial phagocytosis in BV2 cells. (a) The representative confocal z-stack fluorescent image DAPI (blue), Iba1 (green), and Aβ1-42 (red) focused on microglial recruitment and plaque deposition in the hippocampus proper and medial entorhinal cortex between vehicle and 3 months Avn-C oral administered hippocampal tissue sections (scale bar 200 µm). (b) Fluorescent confocal z-stack image from 5xFAD vehicle and three months Avn-C-administered mice hippocampal sections (50 µm, two sections per mouse) fluorescent tag with Iba1 (green) and Aβ42 (red) (scale bar 20 µm). (c) Quantification of the area covered by microglia over the plaque between vehicle and three months Avn-C orally administered mice from early AD stage from hippocampus proper. The ratio is calculated as (area of microglia)/(area of plaque) in the region of interest. Number of plaque analyzed 30/group (n = 4). (d) Confocal fluorescent image of DAPI (blue) and BV-2 microglial cells (green) pre-activated by oAβ1-42 (1.0 µM) for 30 min and continued treatment with oAβ1-42 (1.0 µM) and oAβ1-42 (1.0 µM) + Avn-C (50 µm) for different time points: 3, 6, and 12 h. Phagocytosis was observed from ingested fluorescent carboxylate microsphere (red) (scale bar 20 µm). (e) Quantification of phagocytosis from BV-2 cells treated with oAβ1-42 (1.0 µM) or oAβ1-42 (1.0 µM) + Avn-C (50 µm), analyzed based on the number of microspheres engulfed per cells from 3, 6, and 12 h time duration; 30 cells analyzed per condition. Mean ± S.E.M.s. Differences were analyzed using an unpaired t-test, two-tailed. Statistical significance is indicated as *** p < 0.001, ** p < 0.01.

Article Snippet: Slides were mounted with ProLong Gold Antifade reagent with DAPI (Cell Signaling, Danvers, MA, USA).

Techniques: Two Tailed Test

Journal: Communications Biology

Article Title: NUPR1 protects against hyperPARylation-dependent cell death

doi: 10.1038/s42003-022-03705-1

Figure Lengend Snippet: a OXPHOS metabolism, reflected by oxygen consumption rate (OCR) levels were measured in MiaPaCa-2 cells after 24 h treatments. b MiaPaCa-2 cells were treated with ZZW-115 (1.5 µM) and olaparib (25 µM) for 24 h, then, loaded with MitoTracker Deep-Red FM and, after fixation, stained with DAPI. Flow cytometry analysis were carried out using MitoProbe™ TMRM ( c ), MitoSOX™ Red ( d ), CellROX™ Orange Reagent ( e ) or Fluo-4-AM ( f ) for analysis of the mitochondrial membrane potential, mitochondrial superoxide levels, intracellular ROS levels and the cytosolic calcium concentration, respectively, after 24 h of incubation with the drugs. Data represent mean ± SEM. Two-way, Sidak was used, * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.001. g PLA was performed in MiaPaCa-2 cells in the presence or in the absence of ZZW-115 together or not with 5-FU in the presence or absence of olaparib. Mouse anti-PAR and rabbit anti-Mitofusin2 antibodies were used. A representative experiment is shown ( n = 3). ImageJ was used to count the number of green dots. Mean ± SEM of foci/nucleus are included.

Article Snippet: Finally, samples were mounted using the Prolong Gold antifade reagent with DAPI (Thermon Fisher).

Techniques: Staining, Flow Cytometry, Membrane, Concentration Assay, Incubation

Journal: bioRxiv

Article Title: Preserving endothelial integrity in human saphenous veins during preparation for coronary bypass surgery

doi: 10.1101/2023.08.25.554690

Figure Lengend Snippet: Representative images of immunofluorescent detection of endothelial markers (A) VE-cadherin and (B) eNOS expression (n=7). EC markers are shown in pink while DAPI labeled nuclei are shown in blue. Lower magnification images of the whole vessels contain pseudo-color lines indicating staining positive areas in orange and enlarged images contain orange arrows indicating staining positivity. Pink boxes indicate the amplified image area. Graphs represent the percentage of each vessel lumen determined to be positive for endothelial markers.

Article Snippet: Samples were mounted using FadeStop TM fluorescent mounting reagent with DAPI (GenuIN Biotech #272).

Techniques: Expressing, Labeling, Staining, Amplification