rabbit anti cd41 antibodies  (Proteintech)

Journal: Frontiers in Immunology

Article Title: Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients

doi: 10.3389/fimmu.2023.1285162

Figure Lengend Snippet: Alteration of platelet surface markers in different clinical presentations of dengue patients. (A) Pie chart showing number of dengue patients of different categories as per WHO 2009 classification. (B) Gating strategy used to assess the platelet activation status in dengue patients and healthy subjects. Expression levels of (C) CD41/CD61 and (D) CD62p on platelets of healthy subjects (n=21) and dengue patients (n=65) of different clinical presentations, WS- (n=29), WS+ (n=27) and SD (n=9). HS denotes healthy subjects, DP denotes dengue patients, WS- denotes dengue patients without warning signs, WS+ denotes dengue patients with warning signs, SD represents severe dengue patients and SDG represents severe disease group. Data is represented as mean ± standard error. All the samples were analyzed by non-parametric Mann-Whitney test in which * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001 and **** represents p < 0.0001.

Article Snippet: Membranes were then incubated overnight with primary antibodies - CD63 (Cat no. sc5275 - 1:1000, Santa Cruz), CD9 (Cat no. 13147S - 1:1000, Cell Signaling Technology), CD41 (Cat no. 13807S - 1:1000, Cell Signaling Technology), GAPDH (Cat no. G9545 - 1:5000, Sigma), VE-Cadherin (Cat no. 2158S - 1:1000, Cell Signaling Technology), CD31 (Cat no. 3582S - 1:1000, Cell Signaling Technology), ZO-2 (Cat no. 2847S - 1:1000, Cell Signaling Technology).

Techniques: Activation Assay, Expressing, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients

doi: 10.3389/fimmu.2023.1285162

Figure Lengend Snippet: Platelet count and its correlation with platelet surface markers in dengue patients. (A) Platelet counts in dengue patients with different clinical presentations, WS- (n=29), WS+ (n=27) and SD (n=9). (B) Comparison of levels of CD41/CD61 in healthy subjects (n=21) with dengue patient having platelet count >50,000/μL (n=37) and dengue patient having platelet count <50,000/μL (n=28) respectively. (C) Correlation analyses of thrombocytopenia with CD41/CD61 surface expression on platelets. (D) Comparison of levels of CD62p in healthy subjects (n=21) with dengue patient having platelet count >50,000/μL (n=37) and dengue patient having platelet count <50,000/μL (n=28) respectively. (E) Correlation analyses of thrombocytopenia with CD62p surface expression on platelets. (F) Comparison of platelet counts in dengue patients without (HM-, n=53) and with hemorrhagic manifestations (HM+, n=12). Comparison of levels of (G) CD41/CD61 and (H) CD62p in healthy subjects (n=21) with dengue patients without (HM-, n=53) and with hemorrhagic manifestations (HM+, n=12). HS denotes healthy subjects, WS- denotes dengue patients without warning signs, WS+ denotes dengue patients with warning signs, SD represents severe dengue patients, HM- represents dengue patients without hemorrhagic manifestations and HM+ represents dengue patients with hemorrhagic manifestations. Data is represented as mean ± standard error. Data were analyzed by non-parametric Mann-Whitney test between the two groups. Correlation analyses were performed using two-tailed Spearman’s correlation coefficient test. * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001 and **** represents p < 0.0001.

Article Snippet: Membranes were then incubated overnight with primary antibodies - CD63 (Cat no. sc5275 - 1:1000, Santa Cruz), CD9 (Cat no. 13147S - 1:1000, Cell Signaling Technology), CD41 (Cat no. 13807S - 1:1000, Cell Signaling Technology), GAPDH (Cat no. G9545 - 1:5000, Sigma), VE-Cadherin (Cat no. 2158S - 1:1000, Cell Signaling Technology), CD31 (Cat no. 3582S - 1:1000, Cell Signaling Technology), ZO-2 (Cat no. 2847S - 1:1000, Cell Signaling Technology).

Techniques: Comparison, Expressing, MANN-WHITNEY, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Platelet derived exosomes disrupt endothelial cell monolayer integrity and enhance vascular inflammation in dengue patients

doi: 10.3389/fimmu.2023.1285162

Figure Lengend Snippet: Platelet derived exosomes isolated from dengue patients express CD63 and CD9 proteins. (A) Expression of CD41, CD63 and CD9 proteins in pooled fractions (n=5) of platelets-pellet, microvesicles and exosomes isolated from healthy subjects and each category of dengue patients as shown by western blot. (B) Size distribution of platelet specific exosomes though DLS. Fold change expressions of (C) CD63 and (D) CD9 in dengue patients (n=23), WS- (n=10), WS+ (n=10), SD (n=3), SDG (n=13) compared to healthy subjects (n=11). Comparisons of fold change expression of (E) CD63 and (F) CD9 in exosomes derived from platelets of primary (1° INF, n=5) and secondary (2° INF, n=18) dengue patients with healthy subjects (n=11). HS denotes healthy subjects, DP denotes dengue patients, WS- denotes dengue patients without warning signs, WS+ denotes dengue patients with warning signs, SD represents severe dengue and SDG represents severe disease group. EXO:HS denotes platelet derived exosomes from healthy subjects, EXO:DP denotes platelet derived exosomes from dengue patients, EXO:WS- denotes platelet derived exosomes from dengue patients without warning signs, EXO:WS+ denotes platelet derived exosomes from dengue patients with warning signs, EXO:SD denotes platelet derived exosomes from severe dengue and EXO:SDG denotes platelet derived exosomes from severe disease group. All the samples were analyzed by parametric unpaired t test in which * represents p < 0.05 and ** represents p < 0.01.

Article Snippet: Membranes were then incubated overnight with primary antibodies - CD63 (Cat no. sc5275 - 1:1000, Santa Cruz), CD9 (Cat no. 13147S - 1:1000, Cell Signaling Technology), CD41 (Cat no. 13807S - 1:1000, Cell Signaling Technology), GAPDH (Cat no. G9545 - 1:5000, Sigma), VE-Cadherin (Cat no. 2158S - 1:1000, Cell Signaling Technology), CD31 (Cat no. 3582S - 1:1000, Cell Signaling Technology), ZO-2 (Cat no. 2847S - 1:1000, Cell Signaling Technology).

Techniques: Derivative Assay, Isolation, Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Thrombocyte-derived Dickkopf1 promotes macrophage polarization in the Bleomycin-induced lung injury model

doi: 10.3389/fimmu.2023.1247330

Figure Lengend Snippet: Thrombocyte is a major source of DKK1. (A-C) Upon BLM challenge, C57Bl/6J mice (n = 4 per group) were intraperitoneally injected with an isotype control antibody (Iso Ab) or anti-CD41a antibody (α-CD41 Ab) on day 0. The lungs were harvested on day 2 (A) . DKK1 protein levels were analyzed by IHC and Image J (B) . Representative IHC images are shown (C) . The boxed areas are shown in higher magnification below. DKK1-positive cells are indicated by red arrows. Bar = 20 μm (top and middle panels); 5 μm (bottom panels). (D) Peripheral blood was collected from Dkk1 PKO mice (n = 5) and their Dkk1 fl/fl littermate controls (n = 5). Circulating DKK1 protein levels were analyzed by DKK1 ELISA. (E) Hemavet analysis of inflammatory cells in the peripheral blood was performed from Dkk1 PKO mice (n = 6) and their Dkk1 fl/fl littermate controls (n = 6). Shown are white blood cells (WBC), lymphocytes, monocytes, neutrophils, eosinophils, basophils, red blood cells (RBC), platelets, and mean platelet volume (MPV). A one-way ANOVA analysis with Bonferroni’s post hoc test was performed for (B) . Student’s t-test was performed for (D, E) . ****p < 0.0001, ns, not significant. See also .

Article Snippet: The sections were incubated with 5% BSA and subsequently stained with primary antibody (Goat polyclonal anti-DKK1, AF1096, R&D systems (6 μg/ml); Rabbit polyclonal anti-CD41 PA5-22307, Thermo Fisher Scientific (1:50)).

Techniques: Injection, Enzyme-linked Immunosorbent Assay