Journal: eLife

Article Title: Microglial SIRPα regulates the emergence of CD11c+ microglia and demyelination damage in white matter

doi: 10.7554/elife.42025

Figure Lengend Snippet: Figure 4. Activation of microglia in the brain white matter of CD47 KO mice. (A) Immunofluorescence staining of coronal brain sections prepared from control (WT) or CD47 KO mice at 19 wks of age with antibodies to Iba1 (red) and CD11c (green). Merged images are shown. The boxed areas in the upper panels are shown at higher magnification in the lower panels. fi, fimbria. Scale bars: 100 mm (upper panels), 50 mm (lower panels). (B) Figure 4 continued on next page

Article Snippet: DOI: https://doi.org/10.7554/eLife.42025 19 of 29 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Antibody PE conjugated rat mAb to mouse CD172a (SIRPa) (clone P84) eBioscience (Cat# 12-1721-80) RRID:AB_11149864 FCM (1:100) Antibody PE conjugated rat mAbs to CD68 (clone FA-11) BioLegend (Cat# 137013) RRID:AB_10613469 FCM (1:100) Antibody PE conjugated rat mAb to mouse CD14 (clone Sa14-2) BioLegend (Cat# 123309) RRID:AB_940582 FCM (1:100) Antibody PE conjugated recombinant antibody (Ab) to Dectin-1 (REA154) Miltenyi Biotec (Cat# 130-102-987) RRID:AB_2651541 FCM (1:5) Antibody PE conjugated recombinant Ab to CD47 (REA170) Miltenyi Biotec (Cat# 130-103-108) RRID:AB_2659745 FCM (1:10) Antibody Rat mAb to myelin basic protein (MBP) (clone 12) Merck (Cat# MAB386) RRID:AB_94975 IHC (1:500) Antibody Rabbit pAb to Olig2 Immuno-Biological Laboratories (Gunma, Japan) (Cat# 18953) RRID:AB_1630817 IHC (1:400) Antibody Alexa Fluor 488 goat anti-rabbit IgG Molecular Probes (Cat# A11034) RRID:AB_2576217 IHC (1:200) Antibody Cy3-conjugated AffiniPure Goat anti-rabbit IgG Jackson Immuno Research (Cat# 111-165-144) RRID:AB_2338006 IHC (1:400) Antibody Cy3-conjugated AffiniPure Goat anti-rat IgG Jackson Immuno Research (Cat# 112-165-167) RRID:AB_2338251 IHC (1:200) Antibody Cy3-conjugated AffiniPure Goat anti-mouse IgG Jackson Immuno Research (Cat# 115-165-166) RRID:AB_2338692 IHC (1:400) Antibody Streptavidin, Alexa Fluor 488 conjugate Molecular Probes (Cat# S11223) RRID:AB_2336881 IHC (1:400) Commercial assay or kit Black-Gold II myelin staining kit Merck Cat# AG105 Commercial assay or kit Tyramide Signal Amplification (TSA) Biotin System kit Perkin Elmer Cat# NEL700A001KT Commercial assay or kit RNeasy Mini kit Qiagen Cat# 74106 Commercial assay or kit QuantiTect Reverse Transcription kit Qiagen Cat# 205313 Commercial assay or kit QuantiTect SYBR Green PCR kit Qiagen Cat# 204143 or 24163 Commercial assay or kit GeneChip 3’IVT Express Kit Affymetrix Cat# 901228 or 901229 Commercial assay or kit Ovation Pico WTA system V2 NuGEN Cat# 3302–12/ 60/–A01 Commercial assay or kit Encore Biotin Module NuGEN Cat# 4200–12/ 60/–A01 Chemical compound, drug Tamoxifen Toronto Research Chemicals Inc. Cat# T006000 Chemical compound, drug 4’,6-Diamidino-2phenylindole Nacalai Tesque (Kyoto, Japan) Cat# 11034–56 Continued on next page Sato-Hashimoto et al. eLife 2019;8:e42025.

Techniques: Activation Assay, Immunofluorescence, Staining, Control

Journal: eLife

Article Title: Microglial SIRPα regulates the emergence of CD11c+ microglia and demyelination damage in white matter

doi: 10.7554/elife.42025

Figure Lengend Snippet: Figure 5. Microarray transcriptome analyses of the white matter and the brain mononuclear cells of CD47 KO mice. (A,B) The results of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis with DAVID. Statistically significant (p-value <0.01) KEGG enrichment pathways of up- (A) or downregulated (B) genes in the white matter (optic nerve and optic tract) (upper panels) or in the brain mononuclear cells (lower panels) of CD47 KO mice. Enrichment score is expressed as –Log (p-value). ARVC, Arrhythmogenic right ventricular cardiomyopathy; HCM, Hypertrophic Figure 5 continued on next page

Article Snippet: DOI: https://doi.org/10.7554/eLife.42025 19 of 29 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Antibody PE conjugated rat mAb to mouse CD172a (SIRPa) (clone P84) eBioscience (Cat# 12-1721-80) RRID:AB_11149864 FCM (1:100) Antibody PE conjugated rat mAbs to CD68 (clone FA-11) BioLegend (Cat# 137013) RRID:AB_10613469 FCM (1:100) Antibody PE conjugated rat mAb to mouse CD14 (clone Sa14-2) BioLegend (Cat# 123309) RRID:AB_940582 FCM (1:100) Antibody PE conjugated recombinant antibody (Ab) to Dectin-1 (REA154) Miltenyi Biotec (Cat# 130-102-987) RRID:AB_2651541 FCM (1:5) Antibody PE conjugated recombinant Ab to CD47 (REA170) Miltenyi Biotec (Cat# 130-103-108) RRID:AB_2659745 FCM (1:10) Antibody Rat mAb to myelin basic protein (MBP) (clone 12) Merck (Cat# MAB386) RRID:AB_94975 IHC (1:500) Antibody Rabbit pAb to Olig2 Immuno-Biological Laboratories (Gunma, Japan) (Cat# 18953) RRID:AB_1630817 IHC (1:400) Antibody Alexa Fluor 488 goat anti-rabbit IgG Molecular Probes (Cat# A11034) RRID:AB_2576217 IHC (1:200) Antibody Cy3-conjugated AffiniPure Goat anti-rabbit IgG Jackson Immuno Research (Cat# 111-165-144) RRID:AB_2338006 IHC (1:400) Antibody Cy3-conjugated AffiniPure Goat anti-rat IgG Jackson Immuno Research (Cat# 112-165-167) RRID:AB_2338251 IHC (1:200) Antibody Cy3-conjugated AffiniPure Goat anti-mouse IgG Jackson Immuno Research (Cat# 115-165-166) RRID:AB_2338692 IHC (1:400) Antibody Streptavidin, Alexa Fluor 488 conjugate Molecular Probes (Cat# S11223) RRID:AB_2336881 IHC (1:400) Commercial assay or kit Black-Gold II myelin staining kit Merck Cat# AG105 Commercial assay or kit Tyramide Signal Amplification (TSA) Biotin System kit Perkin Elmer Cat# NEL700A001KT Commercial assay or kit RNeasy Mini kit Qiagen Cat# 74106 Commercial assay or kit QuantiTect Reverse Transcription kit Qiagen Cat# 205313 Commercial assay or kit QuantiTect SYBR Green PCR kit Qiagen Cat# 204143 or 24163 Commercial assay or kit GeneChip 3’IVT Express Kit Affymetrix Cat# 901228 or 901229 Commercial assay or kit Ovation Pico WTA system V2 NuGEN Cat# 3302–12/ 60/–A01 Commercial assay or kit Encore Biotin Module NuGEN Cat# 4200–12/ 60/–A01 Chemical compound, drug Tamoxifen Toronto Research Chemicals Inc. Cat# T006000 Chemical compound, drug 4’,6-Diamidino-2phenylindole Nacalai Tesque (Kyoto, Japan) Cat# 11034–56 Continued on next page Sato-Hashimoto et al. eLife 2019;8:e42025.

Techniques: Microarray

Journal: bioRxiv

Article Title: CD47 as a potential biomarker for the early diagnosis of severe COVID-19

doi: 10.1101/2021.03.01.433404

Figure Lengend Snippet: SARS-CoV-2 infection is associated with increased CD47 levels. A) TF protein abundance in uninfected (control) and SARS-CoV-2-infected (virus) Caco-2 cells (data derived from . P-values were determined by two-sided Student’s t-test. B) CD47 and SARS-CoV-2 N protein levels and virus titres (genomic RNA determined by PCR) in SARS-CoV-2 strain FFM7 (MOI 1)-infected air-liquid interface cultures of primary human bronchial epithelial (HBE) cells and SARS-CoV-2 strain FFM7 (MOI 0.1)-infected Calu-3 cells. Uncropped blots are provided in Suppl. Figure 1. C) CD47 mRNA levels in post mortem samples from COVID-19 patients (data derived from ). P-values were determined by two-sided Student’s t-test.

Article Snippet: Detection occurred by using specific antibodies against CD47 (1:100 dilution, CD47 Antibody, anti-human, Biotin, REAfinityTM, # 130-101-343, Miltenyi Biotec), SARS-CoV-2 N (1:1000 dilution, SARS-CoV-2 Nucleocapsid Antibody, Rabbit MAb, #40143-R019, Sino Biological), and GAPDH (1:1000 dilution, Anti-G3PDH Human Polyclonal Antibody, #2275-PC-100, Trevigen).

Techniques: Infection, Quantitative Proteomics, Control, Virus, Derivative Assay

Journal: bioRxiv

Article Title: CD47 as a potential biomarker for the early diagnosis of severe COVID-19

doi: 10.1101/2021.03.01.433404

Figure Lengend Snippet: Results of the PubMed ( https://pubmed.ncbi.nlm.nih.gov ) literature search for “CD47 aging” (A) and “CD47 hypertension” (B). C) Overview figure of the data derived from the literature searches. Age-related increased CD47 levels may contribute to pathogenic conditions associated with severe COVID-19.

Article Snippet: Detection occurred by using specific antibodies against CD47 (1:100 dilution, CD47 Antibody, anti-human, Biotin, REAfinityTM, # 130-101-343, Miltenyi Biotec), SARS-CoV-2 N (1:1000 dilution, SARS-CoV-2 Nucleocapsid Antibody, Rabbit MAb, #40143-R019, Sino Biological), and GAPDH (1:1000 dilution, Anti-G3PDH Human Polyclonal Antibody, #2275-PC-100, Trevigen).

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: CD47 as a potential biomarker for the early diagnosis of severe COVID-19

doi: 10.1101/2021.03.01.433404

Figure Lengend Snippet: Results of the PubMed ( https://pubmed.ncbi.nlm.nih.gov ) literature search for “CD47 diabetes” (A). B) Overview figure of the data derived from the literature search. Hyperglycaemia- and diabetes-induced increased CD47 levels may contribute to immune escape of SARS-CoV-2-infected cells.

Article Snippet: Detection occurred by using specific antibodies against CD47 (1:100 dilution, CD47 Antibody, anti-human, Biotin, REAfinityTM, # 130-101-343, Miltenyi Biotec), SARS-CoV-2 N (1:1000 dilution, SARS-CoV-2 Nucleocapsid Antibody, Rabbit MAb, #40143-R019, Sino Biological), and GAPDH (1:1000 dilution, Anti-G3PDH Human Polyclonal Antibody, #2275-PC-100, Trevigen).

Techniques: Derivative Assay, Infection

Journal: Advanced Science

Article Title: Overcoming the On‐Target Toxicity in Antibody‐Mediated Therapies via an Indirect Active Targeting Strategy

doi: 10.1002/advs.202206912

Figure Lengend Snippet: Preparation and characterization of anti‐CD47‐PCM@NP. A) Hydrodynamic size and zeta potential of CM vesicles, PLGA cores (NP), PCM@NP, and anti‐CD47‐PCM@NP. Data are means ± SD ( n = 3). B) Colocalization of NP/C6 (green) with DiD‐PCM (red), and the colocalization of FITC‐antibody (green) with DiD‐PCM@NP (red), both assessed by confocal laser scanning microscope (CLSM) (scale bar = 5 µm). C) Transmission electron micrographs of (a) NP, (b) CM vesicle, (c) PCM@NP, (d) Anti‐CD47‐PCM@NP, and (e) multiple anti‐CD47‐PCM@NP. All scale bars = 100 nm. D) SDS‐PAGE protein analysis of NP, PCM@NP, CM vesicles, and cancer cell lysate. Samples were tested at equal protein concentrations. CD47 protein and membrane‐specific protein on the cancer cell membrane were efficiently retained on the extracted membrane vesicles and the PCM@NP, detected by western blot. E) Determination of the antibody labeled by PE loaded on the surface of anti‐CD47‐PCM@NP by flow nanoanalyzer. F) The binding affinity of the antibody to the CM vesicles by surface plasmon resonance (SPR).

Article Snippet: Streptavidin Conjugation Kit‐Lightning‐Link (Abcam, ab102921). siRNAs (sense 5’‐3’ CACCGAAGAAAUGUUUGUGAATT; sense 5’‐3’ CCAUACGAAUAAGAGAAUCAUTT) were purchased from Shanghai Sangon Biotechnology Co., Ltd. Quantum R‐PE MESF Medium Level (FCSC827B) was from Bio‐Rad Laboratories, Inc. PE anti‐human/mouse/rat CD47 antibody (E‐AB‐F1016D) was purchased from Elabscience (Wuhan, China).

Techniques: Zeta Potential Analyzer, Laser-Scanning Microscopy, Transmission Assay, SDS Page, Membrane, Western Blot, Labeling, Binding Assay, SPR Assay

Journal: Advanced Science

Article Title: Overcoming the On‐Target Toxicity in Antibody‐Mediated Therapies via an Indirect Active Targeting Strategy

doi: 10.1002/advs.202206912

Figure Lengend Snippet: Anti‐CD47‐PCM@NP effectively distinguished target cells from CD47‐expressing nontarget cells in vitro through indirect active targeting. A) Anti‐CD47‐PCM@NP avoided the blocking of CD47 on RBC and subsequent phagocytosis thus circumvented the on‐target toxicity of free anti‐CD47 towards RBC through the INTACT strategy. Scale bar = 50 µm. B,C) PCM@NP and anti‐CD47‐PCM@NP efficiently escaped the capture by B) macrophages with enhanced and parallel cellular uptake by C) target 4T1 cells, measured by flow cytometry. The antibody selectively dissociated from the carrier PCM@NP at the presence of 4T1 cells with high expression of D) CD47 in contrast to E) CD47 −/− 4T1 cells, shown by colocalization images and distribution map of PE‐antibody (red) and PCM@NP/C6 (green). Scale bar = 20 µm (multi‐cell images), 2 µm (single‐cell images). F) The schematic diagram of the microfluidic device. The tumor cells were cultured in the cavity of the microfluidic chip till adherence, and then exposed to flowing anti‐CD47 or anti‐CD47‐PCM@NP, and fluorescent images were captured at predetermined time points. G) Free anti‐CD47 sufficiently bound to the surface of 4T1 cells with high expression of CD47 (a). The antibody dissociated from PCM@NP at the presence of 4T1 cells (b), in contrast with CD47 −/− 4T1 group (c) (antibody labeled with FITC, green. PCM@NP labeled with DiD, red). Scale bar = 10 µm. Original movies are shown in Movie S1 (Supporting Information) (a), Movie S2 (Supporting Information) (b), and Movie S3 (Supporting Information) (c), respectively. Data are presented as mean ± SD ( n = 3). (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS represents non‐significance).

Article Snippet: Streptavidin Conjugation Kit‐Lightning‐Link (Abcam, ab102921). siRNAs (sense 5’‐3’ CACCGAAGAAAUGUUUGUGAATT; sense 5’‐3’ CCAUACGAAUAAGAGAAUCAUTT) were purchased from Shanghai Sangon Biotechnology Co., Ltd. Quantum R‐PE MESF Medium Level (FCSC827B) was from Bio‐Rad Laboratories, Inc. PE anti‐human/mouse/rat CD47 antibody (E‐AB‐F1016D) was purchased from Elabscience (Wuhan, China).

Techniques: Expressing, In Vitro, Blocking Assay, Flow Cytometry, Cell Culture, Labeling

Journal: Advanced Science

Article Title: Overcoming the On‐Target Toxicity in Antibody‐Mediated Therapies via an Indirect Active Targeting Strategy

doi: 10.1002/advs.202206912

Figure Lengend Snippet: Evaluation of the biological functions of anti‐CD47‐PCM@NP in vivo. A) In vivo and ex vivo targeting ability of anti‐CD47‐PCM@NP and anti‐CD47 in tumor‐bearing mice models determined by live imaging. B) The semiquantitative analysis of the ratio of fluorescence intensity (tumor/liver) of ex vivo imaging. C) In vivo biodistribution of coumarin 6 (C6)‐labeled formulations in tumor‐bearing mice models at (a) 2 h, (b) 4 h, (c) 8 h, and (d) 12 h after i.v. injection of NP/C6, PCM@NP/C6 and anti‐CD47‐PCM@NP/C6. Data are presented as mean ± SD ( n = 3) (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS represents non‐significance).

Article Snippet: Streptavidin Conjugation Kit‐Lightning‐Link (Abcam, ab102921). siRNAs (sense 5’‐3’ CACCGAAGAAAUGUUUGUGAATT; sense 5’‐3’ CCAUACGAAUAAGAGAAUCAUTT) were purchased from Shanghai Sangon Biotechnology Co., Ltd. Quantum R‐PE MESF Medium Level (FCSC827B) was from Bio‐Rad Laboratories, Inc. PE anti‐human/mouse/rat CD47 antibody (E‐AB‐F1016D) was purchased from Elabscience (Wuhan, China).

Techniques: In Vivo, Ex Vivo, Imaging, Fluorescence, Labeling, Injection

Journal: Advanced Science

Article Title: Overcoming the On‐Target Toxicity in Antibody‐Mediated Therapies via an Indirect Active Targeting Strategy

doi: 10.1002/advs.202206912

Figure Lengend Snippet: Antitumor efficacy of anti‐CD47‐PCM@NP and mechanistic investigation by CyTOF analysis. A) Representative images and phagocytic index of C57BL/6 bone marrow‐derived macrophages (BMDM) phagocytosing tumor cells following treatment with PCM@NP, anti‐CD47, and anti‐CD47‐PCM@NP. Scale bar = 50 µm. B) Timeline of the anti‐tumor efficacy study on tumor‐bearing mice (red arrows indicate intravenous administrations), and average tumor growth curves and picture of tumor tissues after the treatment. C) Individual tumor growth curves in each group. D) viSNE plot of intratumoral cells in tumor tissues after treatment with saline, PCM@NP, anti‐CD47, anti‐CD47‐PCM@NP and all groups merged. E) Heat map of the surface molecule and functional molecule expression of different subsets of immune cells in tumor tissues from all groups merged. F) tSNE visualization of all samples with the expression of CD4 and CD8a respectively. G) Percentage of cells in each cluster after treatment from each group. Data represented as mean ± SD ( n = 6). (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS represents nonsignificance).

Article Snippet: Streptavidin Conjugation Kit‐Lightning‐Link (Abcam, ab102921). siRNAs (sense 5’‐3’ CACCGAAGAAAUGUUUGUGAATT; sense 5’‐3’ CCAUACGAAUAAGAGAAUCAUTT) were purchased from Shanghai Sangon Biotechnology Co., Ltd. Quantum R‐PE MESF Medium Level (FCSC827B) was from Bio‐Rad Laboratories, Inc. PE anti‐human/mouse/rat CD47 antibody (E‐AB‐F1016D) was purchased from Elabscience (Wuhan, China).

Techniques: Derivative Assay, Saline, Functional Assay, Expressing

Journal: Advanced Science

Article Title: Overcoming the On‐Target Toxicity in Antibody‐Mediated Therapies via an Indirect Active Targeting Strategy

doi: 10.1002/advs.202206912

Figure Lengend Snippet: The INTACT strategy efficiently delivers antibodies to tumors with reduced in vivo toxicity. A) Anti‐CD47‐PCM@NP exhibited no significant influence on red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), and platelet (PLT). Data represented as mean ± SD ( n = 3). B–D) Anti‐CD47‐PCM@NP relieved the occurrence of fungal infection during antitumor treatment. B) Experimental timeline and treatments in tumor‐bearing mice (arrows indicate intravenous administrations). At day 14, mice were infected with C. albicans via tail vein injection. C) Colony‐forming units (CFU) on day 7 in the kidneys of infected mouse models ( n = 6). D) The survival rates of infected mice with different treatments ( n = 12). Data represented as mean ± SD. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS represents non‐significance).

Article Snippet: Streptavidin Conjugation Kit‐Lightning‐Link (Abcam, ab102921). siRNAs (sense 5’‐3’ CACCGAAGAAAUGUUUGUGAATT; sense 5’‐3’ CCAUACGAAUAAGAGAAUCAUTT) were purchased from Shanghai Sangon Biotechnology Co., Ltd. Quantum R‐PE MESF Medium Level (FCSC827B) was from Bio‐Rad Laboratories, Inc. PE anti‐human/mouse/rat CD47 antibody (E‐AB‐F1016D) was purchased from Elabscience (Wuhan, China).

Techniques: In Vivo, Infection, Injection

Journal: Advanced Science

Article Title: Overcoming the On‐Target Toxicity in Antibody‐Mediated Therapies via an Indirect Active Targeting Strategy

doi: 10.1002/advs.202206912

Figure Lengend Snippet: The INTACT strategy is adaptive to multiple antibody‐based systems. A) Relative tumor volume growth with anti‐CD47‐PCM@NP/PTX treatment ( n = 6). B–G) The INTACT therapy refined the targeting precision of ADC. B) The diagram of ADC construction: Anti‐CD47 was modified with streptavidin and conjugated with DM1 via the crosslinker BMCC‐biotin. C) The conjugation of ADC was confirmed with SDS‐PAGE. D) Experimental timeline for the anti‐tumor efficacy study and hematology assessments of ADC‐PCM@NP (red arrows indicate intravenous administrations). E) Average tumor growth curves, and picture of the tumor tissues after the treatment ( n = 6). F) Individual tumor growth curves in each group ( n = 6). G) Hematology assessments of red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), and platelet (PLT) ( n = 3). Data represented as mean ± SD. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS represents non‐significance).

Article Snippet: Streptavidin Conjugation Kit‐Lightning‐Link (Abcam, ab102921). siRNAs (sense 5’‐3’ CACCGAAGAAAUGUUUGUGAATT; sense 5’‐3’ CCAUACGAAUAAGAGAAUCAUTT) were purchased from Shanghai Sangon Biotechnology Co., Ltd. Quantum R‐PE MESF Medium Level (FCSC827B) was from Bio‐Rad Laboratories, Inc. PE anti‐human/mouse/rat CD47 antibody (E‐AB‐F1016D) was purchased from Elabscience (Wuhan, China).

Techniques: Modification, Conjugation Assay, SDS Page

Journal: Frontiers in Oncology

Article Title: Spatially resolved transcriptomics revealed local invasion-related genes in colorectal cancer

doi: 10.3389/fonc.2023.1089090

Figure Lengend Snippet: Cell differentiation trajectories in CRC obtained by pseudotime analysis. (A–D) tSNE map shows the results of the dimensionality reduction and clustering analysis of S112 (A) , S115 (B) , S114 (C) , and 927 (D) (up). Results of pseudotime cell trajectory in S112 (A) , S115 (B) , S114 (C) , and S927 (D) (down). (E) Twelve genes were screened by invasive modules. (F) The relationship of STC1 expression level and cancer stage/progression-free survival in colon cancer (up) and rectal cancer (down) from TCGA database. (G) The relationship of CES1 expression level and cancer stage in colon cancer (up) and rectal cancer (down) from TCGA database. (H) Immunohistochemical staining showed the expression of AKR1B1(left panel), STC1(middle panel), and CD47(right panel) in Mucosa and cancer(up) and Invasive margin(down) (n=45). The scale bars on the lower right are 100 µm. *P < 0.05, **P < 0.01 and ***P < 0.001. NS, not significant difference.

Article Snippet: Antibodies used include STC1 (1:50; Proteintech, China), RPL5(1:800; Proteintech, China), AKR1B1(1:50; BOSTER, China), HLA-A(1:100; BOSTER, China) and CD47(1:400; BOSTER, China).

Techniques: Cell Differentiation, Expressing, Immunohistochemical staining, Staining

Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Article Title: Anti-CD47 Monoclonal Antibody Therapy Reduces Ischemia-Reperfusion Injury of Renal Allografts in a Porcine Model of Donation after Cardiac Death

doi: 10.1111/ajt.14567

Figure Lengend Snippet: (A) The CD47mAb was strongly bound to the renal tissue immediately after flushing. (B) CD47mAb binding to glomeruli could be detected on day 5 after transplant. (These staining were performed on grafts from different animals.)

Article Snippet: Just prior to implantation, the grafts were flushed either with control (n=4) or a humanized CD47mAb (clone anti-CD47 649, Tioma Therapeutics, Inc, St. Louis, n=4) at a dosage of 10 mg per 250–300 g kidney.

Techniques: Binding Assay, Staining

Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Article Title: Anti-CD47 Monoclonal Antibody Therapy Reduces Ischemia-Reperfusion Injury of Renal Allografts in a Porcine Model of Donation after Cardiac Death

doi: 10.1111/ajt.14567

Figure Lengend Snippet: Graft blood flow prior (0s) and post reperfusions at 1s, 15s, 30s, and 60s were studied using an in vivo imaging system (n=2/group). (A) Representative perfusion using ICG fluorescence imaging are shown at time points 0, 16 and 60 seconds after release of clamps (see Figure S1 for additional images). (B) The fluorescence density values of the region of interest (ROI) were extracted to evaluate reperfusion characteristics of the grafts. CD47mAb-treated grafts had more uniform and greater average tissue perfusion as compared to control organs. (C) The repeated measure two-way ANOVA shows that Time (p<0.001), Treatment (p=0.002), and Interaction (p<0.001) terms are all significant, indicating florescent density differed by treatment and over time.

Article Snippet: Just prior to implantation, the grafts were flushed either with control (n=4) or a humanized CD47mAb (clone anti-CD47 649, Tioma Therapeutics, Inc, St. Louis, n=4) at a dosage of 10 mg per 250–300 g kidney.

Techniques: In Vivo Imaging, Fluorescence, Imaging

Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Article Title: Anti-CD47 Monoclonal Antibody Therapy Reduces Ischemia-Reperfusion Injury of Renal Allografts in a Porcine Model of Donation after Cardiac Death

doi: 10.1111/ajt.14567

Figure Lengend Snippet: (A) Serum creatinine, blood urea nitrogen (BUN), and phosphorus were significantly decreased, while the calcium levels were significantly increased in the CD47mAb-treated group than that in the control group. (B) Histological study of pig kidney grafts at day 5 post-KTx indicated that CD47mAb treated grafts had significantly less evidence of acute tubular injury (ATI) compared with the control group. (C) Animals with CD47mAb treated grafts had significantly earlier urination than that in control (n=4/group, CD47mAb treatment versus control, *p<0.05 or **p<0.01, respectively).

Article Snippet: Just prior to implantation, the grafts were flushed either with control (n=4) or a humanized CD47mAb (clone anti-CD47 649, Tioma Therapeutics, Inc, St. Louis, n=4) at a dosage of 10 mg per 250–300 g kidney.

Techniques:

Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Article Title: Anti-CD47 Monoclonal Antibody Therapy Reduces Ischemia-Reperfusion Injury of Renal Allografts in a Porcine Model of Donation after Cardiac Death

doi: 10.1111/ajt.14567

Figure Lengend Snippet: (A) Revealed by qRT-PCR, CD47mAb-treated organs showed significantly less expression in each of these oxidative injury-related genes of superoxide dismutase-1 (sod-1), glutathione peroxidase-1 (gpx-1), thioredoxin (txn), except heme oxygenase-1 (hmox-1) compared to control. (B) Immunofluorescence staining revealed a decreased expression of Voltage-dependent anion-selective channel protein 1 (VDAC-1) and poly (ADP-ribose) polymerase (PARP) in the CD47mAb-treated tissues than in control. (C) Western blotting and (D) densitometry analysis also support that the protein levels of VDAC-1 and PARP are decreased in the grafts treated with CD47mAb compared with control (n=4/group, *p<0.05, ** p<0.01, or *** p<0.001, respectively).

Article Snippet: Just prior to implantation, the grafts were flushed either with control (n=4) or a humanized CD47mAb (clone anti-CD47 649, Tioma Therapeutics, Inc, St. Louis, n=4) at a dosage of 10 mg per 250–300 g kidney.

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Western Blot

Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Article Title: Anti-CD47 Monoclonal Antibody Therapy Reduces Ischemia-Reperfusion Injury of Renal Allografts in a Porcine Model of Donation after Cardiac Death

doi: 10.1111/ajt.14567

Figure Lengend Snippet: (A) qRT-PCR showed the relative mRNA levels of il-2, il-6, tgf-b, and inf-g were significantly lower in the CD47mAb treated renal allografts than that in control at day 5 after kidney transplantation. (B) Immunofluorescence staining indicated the CD4+ and CD8+ cells infiltration were significantly decreased in the CD47mAb treated renal allografts than that in the control (n=4/group, *p<0.05 or ** p<0.01, respectively).

Article Snippet: Just prior to implantation, the grafts were flushed either with control (n=4) or a humanized CD47mAb (clone anti-CD47 649, Tioma Therapeutics, Inc, St. Louis, n=4) at a dosage of 10 mg per 250–300 g kidney.

Techniques: Quantitative RT-PCR, Transplantation Assay, Immunofluorescence, Staining

Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Article Title: Anti-CD47 Monoclonal Antibody Therapy Reduces Ischemia-Reperfusion Injury of Renal Allografts in a Porcine Model of Donation after Cardiac Death

doi: 10.1111/ajt.14567

Figure Lengend Snippet: (A) Immunofluorescence staining showed a reduction of matrix metalloproteinase-2 (MMP-2) and MMP-9 in CD47mAb treated renal allografts than that in control. Western blotting (B) and densitometry analysis (C) confirmed the decreasing expression of MMP-2 and MMP-9 in renal grafts treated with CD47mAb compared to control (n=4/group, *p<0.05 or ** p<0.01, respectively).

Article Snippet: Just prior to implantation, the grafts were flushed either with control (n=4) or a humanized CD47mAb (clone anti-CD47 649, Tioma Therapeutics, Inc, St. Louis, n=4) at a dosage of 10 mg per 250–300 g kidney.

Techniques: Immunofluorescence, Staining, Western Blot, Expressing

Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Article Title: Anti-CD47 Monoclonal Antibody Therapy Reduces Ischemia-Reperfusion Injury of Renal Allografts in a Porcine Model of Donation after Cardiac Death

doi: 10.1111/ajt.14567

Figure Lengend Snippet: (A) Immunofluorescence staining showed a decrease of Bcl-2-associated X protein (BAX) and Caspase-3 in the CD47mAb treated renal allografts compared to control. Western blotting (B) and densitometry analysis (C) confirmed the decreasing expression of BAX and Caspase-3 in renal grafts treated with CD47mAb than that in control (n=4/group, *p<0.05 or ** p<0.01, respectively).

Article Snippet: Just prior to implantation, the grafts were flushed either with control (n=4) or a humanized CD47mAb (clone anti-CD47 649, Tioma Therapeutics, Inc, St. Louis, n=4) at a dosage of 10 mg per 250–300 g kidney.

Techniques: Immunofluorescence, Staining, Western Blot, Expressing