Journal: Scientific Reports

Article Title: Low-dose acetaminophen induces early disruption of cell-cell tight junctions in human hepatic cells and mouse liver

doi: 10.1038/srep37541

Figure Lengend Snippet: Histograms representing relative expression of integrins (CD49a-f) and CD29 in HepaRG cells following challenge with different concentrations of APAP. Though the expression of most integrins was modulated over the period of exposure to APAP, only modulation of CD49c was directly associated with increasing APAP concentration. Untreated HepaRG cells contained high and low CD49c expressing populations. 24 hours after treatment, increasing APAP concentration was directly associated with increased percentage of cells in the low expressing population, and a reciprocal decrease in the high expressing population. The level of expression (MFI) was unchanged in the CD49c low population, whereas the level of expression by cells in the high population increased directly with APAP concentration. Detailed flow cytometric data is shown in .

Article Snippet: To assess correlation of z-alpha with modulation of expression of integrins, adhesion molecules and other markers in response to APAP hepatotoxicity, remaining adherent HepaRG cells were recovered using TrypLETM cell-dissociation reagent (Life Technologies), and stained using combinations of directly conjugated monoclonal antibodies: CD29-BV510 (563513), CD49f-BV421 (5625820, CD49d-FITC (580840), CD49c-PE, CD166-PE (559263) (all BD Biosciences, Oxford, UK); CD49a-APC-Vio770 (130-101-406), CD44-APC-Vio770 (130-099-149), CD49b-PE-Vio770 (130-100-328), CD90-PE-Vio770 (130-099-295), CD49e-APC (130-097-221), (all Miltenyi Biotec, Surrey, UK); CD13-BV421 (301716; Biolegend), CD54-FITC (mhcd5401; Caltag, Buckingham, UK).

Techniques: Expressing, Concentration Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Co-targeting CD47 and VEGF elicited potent anti-tumor effects in gastric cancer

doi: 10.1007/s00262-024-03667-9

Figure Lengend Snippet: Targeting CD47 significantly inhibited the growth of gastric cancer in Hu-PDX models. A In the Hu-PDX1 model, tumor volume was measured twice a week and presented as mean ± SD. After treatment with SIRPα-Fc for 4 weeks, tumor weight was presented. ( n = 6 for control group and n = 8 for SIRPα-Fc group, * P < 0.05, ** P < 0.01) B Each line represented the tumor volume from an independent mouse in the Hu-PDX1 model. C In the Hu-PDX2 model, tumor volume was measured twice a week and presented as mean ± SD. After treatment with SIRPα-Fc for 4 weeks, tumor weight was presented. ( n = 6 per group, * P < 0.05, ** P < 0.01). D Each line represented the tumor volume from an independent mouse in the Hu-PDX2 model. E Representative photographs of immunohistochemical staining for CD80, CD163, and CD8 of tumor tissue sections and the number of CD80 + , CD163 + , and CD8 + cells in each group were normalized to the control group. The value of control was set to 1.0. ( * P < 0.05, ** P < 0.01)

Article Snippet: Anti-CD80 monoclonal antibody (66,406-1-Ig, Proteintech), anti-CD163 monoclonal antibody (GB13340, Servicebio), anti-CD8 monoclonal antibody (GB12068, Servicebio), anti-CD31 monoclonal antibody (GB113151, Servicebio), VEGFA Monoclonal antibody (19,003-1-AP, Proteintech), anti-CD47 monoclonal antibody (ab218810, Abcam), carboxyfluorescein diacetate succinimidyl ester (CFDA SE) (C0051, Beyotime), PerCP anti-CD68 (333,813, BioLegend), PE anti-CD11b (101,208, BioLegend), granulocyte–macrophage colony-stimulating factor (GM-CSF) (C003, novoprotein), FITC-labeled anti-CD47 (CC2C6, BioLegend), Human Lymphocyte separation medium (7,111,011, DAKEWE).

Techniques: Control, Immunohistochemical staining, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Co-targeting CD47 and VEGF elicited potent anti-tumor effects in gastric cancer

doi: 10.1007/s00262-024-03667-9

Figure Lengend Snippet: CD47 blockade combined with antiangiogenetic therapy elicited enhanced anti-tumor effect in Hu-PDX models of gastric cancer. A and B In the Hu-PDX1 model, tumor-bearing mice were treated with SIRPα-Fc and/or VEGFR1-Fc for 4 weeks, tumor volume and tumor weight were presented as mean ± SD. Each line represented the value of the tumor volume of a single mouse. C and D In the Hu-PDX2 model, tumor volume and tumor weight were presented after the same treatment in the Hu-PDX1 model. Each line represented the value of the tumor volume of a single mouse. ( n = 6 per group, * P < 0.05, ** P < 0.01). E Representative photographs of immunohistochemical staining for CD80, CD163, CD8, and CD31 of tumor tissue sections and the number of CD80 + , CD163 + , and CD8 + cells and the relative vessel density in each group were normalized to the control group. The value of control was set to 1.0. (* P < 0.05, ** P < 0.01)

Article Snippet: Anti-CD80 monoclonal antibody (66,406-1-Ig, Proteintech), anti-CD163 monoclonal antibody (GB13340, Servicebio), anti-CD8 monoclonal antibody (GB12068, Servicebio), anti-CD31 monoclonal antibody (GB113151, Servicebio), VEGFA Monoclonal antibody (19,003-1-AP, Proteintech), anti-CD47 monoclonal antibody (ab218810, Abcam), carboxyfluorescein diacetate succinimidyl ester (CFDA SE) (C0051, Beyotime), PerCP anti-CD68 (333,813, BioLegend), PE anti-CD11b (101,208, BioLegend), granulocyte–macrophage colony-stimulating factor (GM-CSF) (C003, novoprotein), FITC-labeled anti-CD47 (CC2C6, BioLegend), Human Lymphocyte separation medium (7,111,011, DAKEWE).

Techniques: Immunohistochemical staining, Staining, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Co-targeting CD47 and VEGF elicited potent anti-tumor effects in gastric cancer

doi: 10.1007/s00262-024-03667-9

Figure Lengend Snippet: Bispecific fusion protein SIRPα-VEGFR1 elicited synergetic antitumor effect and prevented gastric cancer recurrence. A and B In the Hu-PDX1 model, tumor volume and tumor weight were measured and the data was presented as mean ± SD after treatment with SIRPα-Fc plus VEGFR1-Fc, and SIRPα-VEGFR1 for 4 weeks. Each line represented the value of the tumor volume of a single mouse. C and D In the Hu-PDX2 model, tumor volume and tumor weight were measured after the same treatment in the Hu-PDX1 model. E The number of CD80 + , CD163 + , and CD8 + cells and the relative vessel density in each group were normalized to the control group. The value of control was set to 1.0. (* P < 0.05, ** P < 0.01). F In the humanized tumor recurrence model, mice were treated with control, SIRPα-Fc + VEGFR1-Fc, SIRPα-VEGFR1 for 2 weeks, and tumor volume was measured. Each line represented the value of the tumor volume of a single mouse. G Survival curves for different treatment groups. (n = 6 per group, * P < 0.05, ** P < 0.01)

Article Snippet: Anti-CD80 monoclonal antibody (66,406-1-Ig, Proteintech), anti-CD163 monoclonal antibody (GB13340, Servicebio), anti-CD8 monoclonal antibody (GB12068, Servicebio), anti-CD31 monoclonal antibody (GB113151, Servicebio), VEGFA Monoclonal antibody (19,003-1-AP, Proteintech), anti-CD47 monoclonal antibody (ab218810, Abcam), carboxyfluorescein diacetate succinimidyl ester (CFDA SE) (C0051, Beyotime), PerCP anti-CD68 (333,813, BioLegend), PE anti-CD11b (101,208, BioLegend), granulocyte–macrophage colony-stimulating factor (GM-CSF) (C003, novoprotein), FITC-labeled anti-CD47 (CC2C6, BioLegend), Human Lymphocyte separation medium (7,111,011, DAKEWE).

Techniques: Control

Journal: Nature communications

Article Title: Macrophage hypoxia signaling regulates cardiac fibrosis via Oncostatin M.

doi: 10.1038/s41467-019-10859-w

Figure Lengend Snippet: Fig. 5 OSM suppresses SMAD signaling. a After pretreatment with the OSM (10 ng per ml) or IL6 (20 ng per ml), C3H/10T1/2 cells were stimulated with TGF-β1 (2.5 ng per ml, 12 h) and the relative expression level of αSMA mRNA was calculated. The one-way ANOVA and Dunnett’s multiple comparisons test were used for the statistical analysis (F (2, 6) = 97.35). b Immunoblot analysis of phospho-ERK1/2, total ERK1/2, phospho-SMAD2 linker (Ser245/ 250/255) and total SMAD2 protein. Thirty minutes after U0126 (20 nM) pretreatment, C3H/10T1/2 cells were stimulated with OSM (10 ng per ml) or IL6 (20 ng per ml) for 30 min. and collected for the analysis. U0126: a selective MEK1/2 inhibitor. c Immunoblot analysis of phospho-SMAD2 C-terminal (Ser465/467) and Lamin A/C. Thirty minutes after OSM (10 ng per ml) pretreatment, C3H/10T1/2 cells were stimulated with TGF-β1 (2.5 ng per ml, 12 h) and collected at indicated time points. d After pretreatment with U0126 (20 μM, 60 min) and OSM (10 ng per ml, 30 min), C3H/10T1/2 cells were stimulated with TGF-β1 (2.5 ng per ml, 12 h) and the relative expression level of αSMA mRNA was calculated. Two-tailed t-test with Welch’s correction was used for the statistical analysis (t = 3.212, df = 3.172). Data show the mean and the standard deviation (error bar) of technical triplicates from a representative experiment (a, d). *p < 0.05

Article Snippet: Recombinant mouse OSM (catalog no. 495-MO-025), mouse IL6 (catalog no. 406-ML-005), and human TGF-β1 (catalog no. 240-B-010) were all purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Western Blot, Two Tailed Test, Standard Deviation

Journal: Neoplasia (New York, N.Y.)

Article Title: Smac mimetics and TRAIL cooperate to induce MLKL-dependent necroptosis in Burkitt's lymphoma cell lines

doi: 10.1016/j.neo.2021.03.003

Figure Lengend Snippet: Smac mimetics induce necroptosis in BL . (A) Cells were treated with BV6 (BL-2: 4 µM, BL-30: 6 µM), AT-406, LCL-161 and Birinapant (2,5 µM), TRAIL (BL-30: 5 ng/mL, BL-2: 15 ng/mL) and zVAD.fmk (20 µM) in the presence and absence of Dabrafenib (BL-30: 5 µM, BL-2: 10 µM), Nec-1s (20 µM) and NSA (1,5 µM) for 24 h (BL-30) or 48 h (BL-2). Cell death was determined by analysis of PI/Hoechst staining and ImageXpress Micro XLS system. (B) Cells were treated for 24 h (BL-30, RAJI, RAMOS, Seraphine, DAUDI) or 48 h (BL-2) with BV6 (BL-2: 4 µM, DAUDI: 5 µM, BL-30: 6 µM, Seraphine: 7 µM, RAJI: 8 µM, RAMOS: 18 µM), TRAIL (BL-30: 5 ng/mL, RAMOS/Seraphine: 10 ng/mL, BL-2, RAJI, DAUDI: 15 ng/mL) and zVAD.fmk (20 µM). mRNA expression of TRAIL-R1 and TRAIL-R2 was analyzed by qRT-PCR and fold change normalized to 28s mRNA is shown. Mean and SEM of 3 independent experiments are shown; * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: LCL-161 was purchased from Novartis and AT-406, Birinapant and SGI-110 (Guadecitabine) were obtained from Selleck Chemicals.

Techniques: Staining, Expressing, Quantitative RT-PCR