Journal: PLOS One

Article Title: Impact of blast exposure on visual pathway: Mechanism exploration and novel diagnostic perspectives

doi: 10.1371/journal.pone.0344993

Figure Lengend Snippet: (A) Schematic diagram of the visual pathway and key observation areas in its intracranial segment. (B) Representative images of immunofluorescence staining for NeuN (green) and NLRP3 (red), Iba-1 (grey) in mouse visual cortex slices, Group information is shown in images. The red triangle marks indicate the co-localization of NLRP3 and NeuN. The scale bars for all fluorescence intensity channels and the merge are set at 50 μm, the magnified inset is 20μm. Quantitative analysis of visual cortex immunofluorescence density in Iba-1 (C) and NLRP3 (G) . (H) Fluorescence intensity of NeuN + NLRP3 + / NeuN + , n = 4. (D, E, F, I, J) Western blot analysis of Iba-1, IL-1β, cleaved Gasdermin D and NLRP3 in visual cortex lysates, the molecular mass is indicated in kilodaltons, n = 3. (K-L) Quantitative analysis of representative fluorescence images of NeuN staining with TUNEL labeling in the peri-contusion region from different groups that information is shown in images, Scale bar = 100 μm, n = 4. (M) Quantitative analysis of visual cortex NeuN + cells, n = 4. Error bars indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.

Article Snippet: Brain sections were subjected to double staining using NeuN antibody and the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay kit (Beyotime, #C1090) to identify DNA damage, following the manufacturer’s protocol.

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot, TUNEL Assay, Labeling

Journal: PLOS One

Article Title: Impact of blast exposure on visual pathway: Mechanism exploration and novel diagnostic perspectives

doi: 10.1371/journal.pone.0344993

Figure Lengend Snippet: (A) Representative images of immunofluorescence staining for NeuN (green), NLRP3 (red), and Iba-1 (grey) in mouse visual cortex slices, as well as group information, are shown in the images. The red triangle marks indicate the co-localization of NLRP3 and NeuN. The scale bars for all fluorescence intensity channels and the merge are set at 50 μm, the magnified inset is 20 μm, n = 4. (B) Quantitative analysis of visual cortex immunofluorescence density in NLRP3, n = 4. (C) Fluorescence intensity of NeuN + NLRP3 + /NeuN + , n = 4. (D) Quantitative analysis of visual cortex immunofluorescence density in Iba-1, n = 4. (E, F, G, H) Western blot analysis of Iba-1, cleaved Gasdermin D and NLRP3 in visual cortex lysates. The molecular mass is indicated in kilodaltons, n = 3. (I, J) Quantitative analysis of representative fluorescence images of NeuN staining with TUNEL labeling in the peri-contusion region from different groups that information is shown in images, Scale bar = 100 μm, n = 4. (K) Quantitative analysis of visual cortex NeuN+ cells. (L) Representative images of immunofluorescence staining for MBP (green) and βIII-Tubulin (red) in mouse optic nerve slices. Group information is shown in images. Scale bar = 20 μm, n = 4. (M) optic nerve MBP density in different groups, n = 4. (N) optic nerve relative fluorescence intensity of MBP + /β III Tubulin + in different groups, n = 4. (O) TEM analysis of the optic nerve between the BE 24 h and BE 28 d groups, red stars indicate axons with obvious demyelination. Error bars indicate mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.

Article Snippet: Brain sections were subjected to double staining using NeuN antibody and the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay kit (Beyotime, #C1090) to identify DNA damage, following the manufacturer’s protocol.

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot, TUNEL Assay, Labeling

Journal: Scientific Reports

Article Title: Effects of leonurine supplementation during in vitro culture on oxidative stress, cell proliferation, apoptosis, and autophagy in bovine embryos

doi: 10.1038/s41598-026-39889-3

Figure Lengend Snippet: Effect of leonurine supplementation during in vitro culture on apoptotic cell number and apoptotic index of bovine embryos. (A) Representative images of TUNEL staining of Day-7 IVEP blastocysts supplemented with 0 (Control) and 20 µM of leonurine. Arrows indicate cells undergoing apoptosis. Apoptotic cell (B), TCC (C), and apoptotic index ratio (D) of blastocysts in the Leo20 ( n = 30) and Con ( n = 30). R = 2. Data are presented as the mean ± SD. The green and orange dots represent the distribution of measurements in the Leo20 and Control groups, respectively. Effect of leonurine on the mRNA expression levels of apoptosis-related genes, BAX (E), CASP3 (F), and BCL2 (G), quantified with RT-qPCR. Data are derived from at least three independent replicates. Abbreviations: BAX, BCL2-related x protein; BCL2, B-cell lymphoma gene-2; CASP3, cysteine aspartic acid protease-3; DAPI, 4’,6-diamidino-2-phenylindole; IVEP, in vitro embryo production; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling; TCC, total cell count. Scale bar = 100 μm.

Article Snippet: The TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) method (In Situ Apoptosis Kit, Green-FITC, E-CK-A320, Elabscience) was utilized to detect cell death and apoptotic index in blastocysts identified after in vitro culture , .

Techniques: In Vitro, TUNEL Assay, Staining, Control, Expressing, Quantitative RT-PCR, Derivative Assay, End Labeling, Cell Characterization

Journal: Scientific Reports

Article Title: Effects of leonurine supplementation during in vitro culture on oxidative stress, cell proliferation, apoptosis, and autophagy in bovine embryos

doi: 10.1038/s41598-026-39889-3

Figure Lengend Snippet: Schematic representation of experimental design. Collection of cumulus-oocyte complexes (COCs) from ovaries obtained from the slaughterhouse, fertilization of oocytes, and in vitro culture of the resulting presumptive zygotes. (A) After fertilization, presumptive zygotes are allocated into different groups (Leo5, 10, 20, 40, 60, and Con) to determine the most effective leonurine dose for embryo development. (B) Following in vitro culture, the potential molecular pathways influenced by leonurine—related to antioxidant activity, apoptosis, cell proliferation, and autophagy—are analyzed using RT-qPCR. (C) To assess embryo quality, various staining procedures are applied, including differential staining, TUNEL, and EdU assays, as well as measurements of ROS, GSH, and LC3B levels.

Article Snippet: The TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) method (In Situ Apoptosis Kit, Green-FITC, E-CK-A320, Elabscience) was utilized to detect cell death and apoptotic index in blastocysts identified after in vitro culture , .

Techniques: In Vitro, Antioxidant Activity Assay, Quantitative RT-PCR, Staining, TUNEL Assay