Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Efficacy of WWQ-131, a highly selective JAK2 inhibitor, in mouse models of myeloproliferative neoplasms.

doi: 10.1016/j.biopha.2022.113884

Figure Lengend Snippet: Fig. 5. WWQ-131 suppressed rhEPO-mediated polycythemia and splenomegaly in BALB/c mice. (A) Reticulocyte count. (B) Hematocrit. (C) Ter119/CD71 eryth roblast population in spleen (top) and bone (down). (D) Spleen weight. (E) Representative photographs of the spleen. (F) p-STAT5 levels in spleen samples. All graphs show mean ± SEM. ###P < 0.001 versus control. *P < 0.05; **P < 0.01; ***P < 0.001 versus model. ^P < 0.05; ^^P < 0.01 versus 120 mg/kg of fedratinib group.

Article Snippet: Anti-JAK2 (Cat No. E-AB-70193), anti-β-ACTIN (Cat No. E-AB-20034), anti-Mouse CD71 (Cat No. E-AB-F1093D) and Ter119 (Cat No. E-AB-F1125E) antibodies were purchased from Elabscience (China).

Techniques: Control

Journal: Oncogene

Article Title: Extracellular vesicle-transported Semaphorin3A promotes vascular permeability in glioblastoma.

doi: 10.1038/onc.2015.317

Figure Lengend Snippet: Figure 3. Tumour-derived EV-transported Sema3A enhances vascular permeability. (a–c) EVs were purified from conditioned medium of GSC#4 that received either non-silencing RNA (siC) or Sema3A-targeting (siS3A), and applied to the left hemisphere of mouse brain. Vehicle (PBS) served as a control. (a) Evans blue extravasation is shown in OCT-included brains. Arrow indicated the site of EV dropping (left hemisphere). (b) Intensity of Evans blue extravasation was quantified in left and right hemispheres, including all brain areas (whole), only cerebrum (crbum) or cerebellum (crbellum). *Po0.05; **Po0.01; ***Po0.001. (c) Confocal analysis of the red blood cell marker Ter119 (red), Claudin5 (Cldn5, green) and Evans Blue (grey) in frozen brain sections. Nucleic acid staining is shown in blue (DAPI). Scale bar: 40 μm. Graph shows the quantification of Ter119 signal intensity from confocal analysis. **Po0.01. (d) Frozen tissue sections from subcutaneous ectopic and intracranial orthotopic GSC#9 xenografts were stained with Pecam (green) and Ter119 (red). Nucleic acid staining is shown in blue (DAPI). Scale bar: 20 μm. (e) Circulating blood EVs were purified from non-injected (non-tum EV, blue) and GSC-xenografted (xeno EV, red) mice and were analysed by flow cytometry for mouse (m) and human (h) CD63. Respective isotype controls (IgG) are shown in grey. Percentage represents the number of positive events for hCD63. (f) Human Sema3A (hSema3A) concentrations were measured by ELISA in circulating EVs purified from sera of non-injected ( −, light orange) and ectopically (ecto.) or orthotopically (ortho.) GSC xenografted (GSC, orange) mice. *Po0.05; **Po0.01. Each panel is representative of at least three independent experiments.

Article Snippet: Antibodies The following antibodies were used: Sema3A (ab23393, Abcam, Paris, France), Alix and Claudin5 (Santa Cruz, Heidelberg, Germany), mouse and human CD63, PECAM (BD Biosciences, Le Pont de Claix, France), Ter119 (R&D Systems, Lille, France), CD9, Hsp70, Rab4, EEA1 (System Biosciences, Ozyme, Saint-Quentin-en-Yvelines, France), Tom20, Kinectin (Cell Signaling, Ozyme), Annexin V, SytoRNA Select and Choleratoxin (Life Technologies).

Techniques: Derivative Assay, Permeability, Control, Marker, Staining, Injection, Cytometry, Enzyme-linked Immunosorbent Assay

Journal: JCI Insight

Article Title: Regulation of bile duct epithelial injury by hepatic CD71 + erythroid cells

doi: 10.1172/jci.insight.135751

Figure Lengend Snippet: (A) H&E staining of liver biopsies at the time of diagnosis of biliary atresia shows variable population of extramedullary cells (absent, scarce, or extensive cell clusters). Scale bar: 20 μM. (B) Immunofluorescence staining of surface markers, CD235a and CD71, identifies erythroblasts among hematopoietic cells. Scale bar: 20 μM. (C) Immunohistochemical staining using anti-CD71 antibody shows similar features in livers of a human neonate and newborn mice. The human liver tissue was obtained at the time of autopsy of a neonate without liver disease, and the mouse livers were obtained from saline- or RRV-injected neonatal mice at 1 day old. Scale bar: 20 μM. (D and E) Quantification of hepatic erythroblasts in response to RRV infection by using flow cytometry after incubation with anti-CD71 and anti-TER119 antibodies at days 1, 3, and 7 after injection of RRV or saline. *P < 0.05, ***P < 0.001. Differences in mean values were statistically analyzed by 2-tailed Student’s t test. n = 3–4/group.

Article Snippet: Fluorochrome-conjugated mouse antibodies were as follows: FITC-conjugated CD71 from eBioscience; APC-conjugated Cd8a and Pan-NK (CD49b) from Miltenyi Biotec; PerCP-conjugated CD3, CD4, CD49b, and anti-NKp46 antibody and BV605-conjugated CD3 and CD45 from BioLegend; and PE-conjugated TER119 from R&D Systems. (catalog FAB1125P).

Techniques: Staining, Biomarker Discovery, Immunofluorescence, Immunohistochemical staining, Saline, Injection, Infection, Flow Cytometry, Incubation

Journal: JCI Insight

Article Title: Regulation of bile duct epithelial injury by hepatic CD71 + erythroid cells

doi: 10.1172/jci.insight.135751

Figure Lengend Snippet: (A and B) Flow cytometry plots and graphical quantification of CD71+Ter119+ erythroblasts after anti-CD71 antibody or rat IgG injection within 12 hours of birth. Livers or extrahepatic bile ducts (EHBD) were harvested at day 1 and day 3 after injection. **P < 0.01, ***P < 0.001, ****P < 0.0001, n = 4 for each group. (C) Weight, jaundice, and survival percentage after the administration of anti-CD71 antibody or isotype (rat IgG) into newborn mice, followed by the i.p. administration of RRV 24 hours later. Two-tailed Student’s t test was used for weight comparison, and χ2 test was used for jaundice percentage. *P < 0.05, ***P < 0.001, ****P < 0.0001; Kaplan-Meier survival curve was analyzed by the log-rank (Mantel-Cox) test; n = 16 for Rat IgG group and n = 21 for anti-CD71 antibody group. (D) H&E staining of liver and EHBD at days 7 and 11 after RRV infection. n = 5 for rat IgG–treated group and n = 14 for anti-CD71 antibody–treated group. PV, portal vein. Scale bar: 20 μM.

Article Snippet: Fluorochrome-conjugated mouse antibodies were as follows: FITC-conjugated CD71 from eBioscience; APC-conjugated Cd8a and Pan-NK (CD49b) from Miltenyi Biotec; PerCP-conjugated CD3, CD4, CD49b, and anti-NKp46 antibody and BV605-conjugated CD3 and CD45 from BioLegend; and PE-conjugated TER119 from R&D Systems. (catalog FAB1125P).

Techniques: Flow Cytometry, Injection, Two Tailed Test, Comparison, Staining, Infection

Journal: JCI Insight

Article Title: Regulation of bile duct epithelial injury by hepatic CD71 + erythroid cells

doi: 10.1172/jci.insight.135751

Figure Lengend Snippet: (A–C) Representative plots and quantification of CD71+Ter119+ cells (A), CD4+ T cells and CD8+ T cells (B), and activation marker NKp46 in CD3–CD49b+ cells and total NK cell counts (C) in livers of RRV-infected mice pretreated with either rat IgG or anti-CD71 antibody. Differences in mean values analyzed by 2-tailed Student’s t test; **P < 0.01; ****P < 0.0001; n = 4/group.

Article Snippet: Fluorochrome-conjugated mouse antibodies were as follows: FITC-conjugated CD71 from eBioscience; APC-conjugated Cd8a and Pan-NK (CD49b) from Miltenyi Biotec; PerCP-conjugated CD3, CD4, CD49b, and anti-NKp46 antibody and BV605-conjugated CD3 and CD45 from BioLegend; and PE-conjugated TER119 from R&D Systems. (catalog FAB1125P).

Techniques: Activation Assay, Marker, Infection

Journal: JCI Insight

Article Title: Regulation of bile duct epithelial injury by hepatic CD71 + erythroid cells

doi: 10.1172/jci.insight.135751

Figure Lengend Snippet: (A) Schematic diagram of the injection timeline for anti-CD71 antibodies or rat IgG isotypes into newborn mice. (B) The quantification of hepatic CD71+Ter119+ cells by flow cytometry shows the prevention of the 3-day surge after RRV infection. Data were analyzed by 1-way ANOVA; ***P < 0.001; ****P < 0.0001; n = 4/group. (C) Jaundice and survival differences in 3 groups. Kaplan-Meier survival curve were analyzed by log-rank (Mantel-Cox) test; *P < 0.05; **P < 0.01; n = 8-16 in each group. (D) CD4+ T cells, CD8+ T cells, and NK cells were quantified by flow cytometry. Data were analyzed by 1-way ANOVA; *P < 0.05; **P < 0.01; ***P < 0.001; n = 3–4/group.

Article Snippet: Fluorochrome-conjugated mouse antibodies were as follows: FITC-conjugated CD71 from eBioscience; APC-conjugated Cd8a and Pan-NK (CD49b) from Miltenyi Biotec; PerCP-conjugated CD3, CD4, CD49b, and anti-NKp46 antibody and BV605-conjugated CD3 and CD45 from BioLegend; and PE-conjugated TER119 from R&D Systems. (catalog FAB1125P).

Techniques: Injection, Flow Cytometry, Infection