Journal: Biosensors

Article Title: Malignant Melanoma-Derived Exosomes Induce Endothelial Damage and Glial Activation on a Human BBB Chip Model

doi: 10.3390/bios12020089

Figure Lengend Snippet: Exosomes derived from A375 cells were more easily absorbed by human brain microvascular endothelial cells. ( A ) Schematic diagram of exosomes purification from tumor cells by differential centrifugation. ( B ) Western blotting showing protein level of exosomal markers (CD63, CD81) in 4 types of tumor cell-derived exosomes ( n = 3). ( C ) Representative TEM images of 4 types of tumor cell-derived exosomes ( n = 2). ( D ) Representative confocal images of hCMEC/D3 cells 24 h after incubation with exosomes (2 μg/mL and 10 μg/mL) derived from 4 different types of tumor cells ( n = 4). Exosomes were labeled by PKH67 (green) fluorescent dye. ( E ) Quantification of PKH67 fluorescence intensity for each group based on ( D ) ( n = 4). Data are presented as mean ± SEM, and are analyzed using a one-way ANOVA with Bonferroni post-test (***: p < 0.001).

Article Snippet: After being blocked with 0.5% skimmed milk powder in TBST buffer containing 0.05% Tween-20, the membranes were probed with the anti-CD63 antibody (Bioworld, BS72936) and anti-CD81 antibody (Proteintech, 66866-1-Ig) at 4 °C overnight, respectively.

Techniques: Derivative Assay, Purification, Centrifugation, Western Blot, Incubation, Labeling, Fluorescence

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Exosomes derived from menstrual blood stromal cells ameliorated premature ovarian insufficiency and granulosa cell apoptosis by regulating SMAD3/AKT/MDM2/P53 pathway via delivery of thrombospondin-1.

doi: 10.1016/j.biopha.2023.115319

Figure Lengend Snippet: Fig. 1. Estrous cycle analysis of the POI rat model in different treatment groups. A. MenSCs after TSP1-siRNA transfection(40 ×, 488 nm, 200 ms); B. The expression of TSP1 in EXOs and MenSCs detected by Western blot (n = 3); C. The gene expression of TSP1 in EXOs detected by RT-PCR (n = 3); D Statistics of cumulative days of different sex cycles in different treatment groups (n = 9); E.Sexual cycle of each rat in different treatment groups. Data was shown as mean ± SD, *p < 0.05， **p < 0.01，***p < 0.001.

Article Snippet: After dewaxed and sodium citrate solution repairing (pH 6.0), ovarian slices of each group were conducted following the instructions of the immunohistochemical kit (KIT-9720, MXB, China) and then incubated with the first antibody at 4 ◦C overnight (TSP1:1:200, proteintech, China; Ki67, 1:200, Wanleibio, China; Caspase 3 1:500, Abmart, China; Caspase 8, 1:500, Abmart, China; PI3K, 1:500, proteintech, China; Phospho-AKT (Ser473), 1:100, Abmart, China; MDM2, 1:200, PTG, China; P53, 1:500, PTG, China; Phospho-SMAD3(Ser425), 1:100, ZENBIO, China).

Techniques: Transfection, Expressing, Western Blot, Gene Expression, Reverse Transcription Polymerase Chain Reaction

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Exosomes derived from menstrual blood stromal cells ameliorated premature ovarian insufficiency and granulosa cell apoptosis by regulating SMAD3/AKT/MDM2/P53 pathway via delivery of thrombospondin-1.

doi: 10.1016/j.biopha.2023.115319

Figure Lengend Snippet: Fig. 2. Ovarian morphology and live births outcomes in different treatment groups. A. Typical photos of the ovaries of rats in different treatment groups; B. His togram of ovarian weight and ovary/body weight ratio; C. TSP1 immunofluorescence and HE staining of paraffin sections of the ovaries in each group; D. Histogram of fluorescence intensity of TSP1; E. Histogram of total follicle counts; F. Histogram of follicle counts in different stages; G. Live birth photos of different groups; H. Comparison of live pups of the EXOs group and EXOsTSP1- group; I. Histogram of total live births in different groups; J. Histogram of the body weight of live births in different groups. Data was shown as mean±SD, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: After dewaxed and sodium citrate solution repairing (pH 6.0), ovarian slices of each group were conducted following the instructions of the immunohistochemical kit (KIT-9720, MXB, China) and then incubated with the first antibody at 4 ◦C overnight (TSP1:1:200, proteintech, China; Ki67, 1:200, Wanleibio, China; Caspase 3 1:500, Abmart, China; Caspase 8, 1:500, Abmart, China; PI3K, 1:500, proteintech, China; Phospho-AKT (Ser473), 1:100, Abmart, China; MDM2, 1:200, PTG, China; P53, 1:500, PTG, China; Phospho-SMAD3(Ser425), 1:100, ZENBIO, China).

Techniques: Immunofluorescence, Staining, Fluorescence, Comparison

Journal: PLOS ONE

Article Title: Identification and validation of TSPAN13 as a novel temozolomide resistance-related gene prognostic biomarker in glioblastoma

doi: 10.1371/journal.pone.0316552

Figure Lengend Snippet: (a) Representative pictures of IHC staining in our glioma tissue microarray cohort (scale bars of f = 200 μm). (b) TSPAN13 protein expression in the glioma tissue microarray. (c). Representative pictures of IHC staining in surgical removal samples (scale bars of g = 100 μm). (d) The expression levels of TSPAN13 in glioma tissues of different grades in the TCGA database. (e) K-M curves of TSPAN13 for GBM based on the TCGA-GBM cohorts. (f) K-M curves of TSPAN13 for GBM based on the Gravendeel cohorts. (g, h) TSPAN13 knockdown efficiency was detected using qRT-PCR and Western blotting in U251 and U87 cell lines. (*p < 0.05, **p < 0.01, ***p < 0.001, ns = not significant).

Article Snippet: The sections were then incubated overnight with primary antibodies (anti- TSPAN13, 18974-1-AP, ProteinTech, China) and then with horseradish peroxidase (HRP)-conjugated secondary antibodies.

Techniques: Immunohistochemistry, Microarray, Expressing, Knockdown, Quantitative RT-PCR, Western Blot

Journal: PLOS ONE

Article Title: Identification and validation of TSPAN13 as a novel temozolomide resistance-related gene prognostic biomarker in glioblastoma

doi: 10.1371/journal.pone.0316552

Figure Lengend Snippet: (a, b) Cell proliferation of U87 and U251 transfected with TSPAN13 siRNA and control was examined by CCK8 assay. (c) Representative pictures of EdU assays in U251 and U87 cell lines. (bar = 50 μm) (d) Statistical results of EdU assays in U251 and U87 cell lines. (e) Representative pictures of transwell and wound healing assays in U251 and U87 cell lines. (f, g) Statistical results of transwell and wound healing assays in U251 and U87 cell lines. (h, i) The GSEA of MAPK and JAK-STAT signalling pathway based on TSPAN13-associated genes. (j) Western blotting analysis showing the expression level of Erk1/2, p-Erk/2, JAK2, p-JAK2, STAT3 and p-STAT3 in TSPAN13 knockdown U87 and U251 cell lines. (*p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: The sections were then incubated overnight with primary antibodies (anti- TSPAN13, 18974-1-AP, ProteinTech, China) and then with horseradish peroxidase (HRP)-conjugated secondary antibodies.

Techniques: Transfection, Control, CCK-8 Assay, Western Blot, Expressing, Knockdown

Journal: PLOS ONE

Article Title: Identification and validation of TSPAN13 as a novel temozolomide resistance-related gene prognostic biomarker in glioblastoma

doi: 10.1371/journal.pone.0316552

Figure Lengend Snippet: (a, b) Cell viability of U87 and U251 treated with TMZ transfected with TSPAN13 siRNA and control was examined by CCK8 assay. (c-f) The expression of γ-H2A.X in U87 and U251 cells transfected with siRNA or treated with TMZ was detected by western blotting and immunofluorescence assay. (g) Representative HE-stained brain sections from tumor-bearing mice in each group of the animal model. (h). Weights of mice are recorded in each group of the animal model. (i). Kaplan–Meier survival of mice in each group. (*p < 0.05, **p < 0.01, ***p < 0.001, ns = not significant).

Article Snippet: The sections were then incubated overnight with primary antibodies (anti- TSPAN13, 18974-1-AP, ProteinTech, China) and then with horseradish peroxidase (HRP)-conjugated secondary antibodies.

Techniques: Transfection, Control, CCK-8 Assay, Expressing, Western Blot, Immunofluorescence, Staining, Animal Model