Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet: FGFR4 is highly expressed in RMS and other cancers, with low expression in healthy tissue (A) High expression of FGFR4 mRNA is found in both FP-RMS and FN-RMS tumors and cell lines compared with other pediatric cancers and healthy tissues. Expression levels measured as FPKM (fragments per kilobase of transcript per million mapped reads) for FGFR4 are summarized in violin plots with medians and quartiles. ASPS, alveolar soft part sarcoma; CCSK, clear cell sarcoma of the kidney; DSRCT, desmoplastic small round cell tumor; EWS, Ewing sarcoma; HBL, hepatoblastoma; ML, melanoma; NB, neuroblastoma; OS, osteosarcoma; SS, synovial sarcoma; UDS, undifferentiated sarcoma; WT, Wilms tumor; YST, yolk sac tumor). (B) FGFR4 mRNA expression in TCGA data shows highest expression in liver hepatocellular carcinoma (LIHC), cholangiocarcinoma (CHOL), and individual tumors of other types. Abbreviations are as per TCGA ( https://gdc.cancer.gov/resources-tcga-users/tcga-code-tables/tcga-study-abbreviations ). (C) Representative images of immunohistochemistry (IHC) for FGFR4 show high expression in RMS, with minimum or no expression in healthy organs. H score displayed in bottom right corner, and scale bars, 200 μm. (D) Summary of membrane-staining H score of FGFR4 IHC of RMS and healthy tissues. Values represent mean ± SEM (error bars). (E) Representative flow cytometry plots show differential levels of FGFR4 expression on FP-RMS or FN-RMS cell lines. Mean fluorescence intensity (MFI) of FGFR4 on indicated RMS cells is listed in the right table, stained with phycoerythrin (PE)-conjugated anti-human FGFR4 antibody or mouse IgG1 isotype control.

Article Snippet: 3A11 is a mouse IgG that was developed from mice immunized with human FGFR4 Fc (R&D).

Techniques: Expressing, Wilms Tumor Assay, Immunohistochemistry, Membrane, Staining, Flow Cytometry, Fluorescence

Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet: Development and characterization of a specific FGFR4 binder from the monoclonal murine antibody 3A11 (A) Structure of the mouse anti-FGFR4 mouse monoclonal antibody (mAb) 3A11. (B) Representative flow cytometry plots show the specificity of 3A11 mAb by staining with FGFR4 + cell lines, FGFR4 KO RH30, or FGFR4 − cell 7250. Mouse IgG (msIgG) is used as isotype control for this mouse antibody 3A11. (C) Structure of anti-FGFR4 antibody 3A11 in scFvFc format fused to the human IgG1 Fc region. (D) Representative flow cytometry plots show 3A11 scFvFc chimeric antibody specifically binds to FGFR4 + cell lines RH30, RH4, and RMS559 but not to the RH30 FGFR4-KO, the RH4 FGFR4-KO, or fibroblast 7250 lines. Human IgG (huIgG) as an isotype control for 3A11 scFvFc. MFI fold change shown as orange font calculated by formula [ M F I ( s c F v F c ) − M F I ( h u I g G i s o t y p e ) ] / M F I ( h u I g G i s o t y p e ) . (E) Binding avidity of FGFR4 scFvFc, using 2-to-1 binding model and global fitting analysis, demonstrates the dissociation constant (K D ) of 3A11 scFvFc against FGFR4 ECD is 4.17 nM. (F) ELISA shows 3A11 scFvFc only recognizes human FGFR4 but not human FGFR1–3 or mouse FGFR4. (G) Flow cytometry using 3A11 scFvFc shows FGFR4 is expressed in several RMS cell lines at various levels with higher expression in FP-RMS compared with in FN-RMS. MFI of 3A11 scFvFc or isotype control huIgG staining on above cells is shown in the table on the right.

Article Snippet: 3A11 is a mouse IgG that was developed from mice immunized with human FGFR4 Fc (R&D).

Techniques: Flow Cytometry, Staining, Binding Assay, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet:

Article Snippet: 3A11 is a mouse IgG that was developed from mice immunized with human FGFR4 Fc (R&D).

Techniques: Purification, Recombinant, Labeling, Virus, Microarray, Cytokine Assay, Transfection, Chromatin Immunoprecipitation, ChIP-sequencing, Plasmid Preparation, Software, Imaging

Journal: Frontiers in Pharmacology

Article Title: A senescence-associated signature refines the classification of different modification patterns and characterization of tumor immune microenvironment infiltration in triple-negative breast cancer

doi: 10.3389/fphar.2023.1191910

Figure Lengend Snippet: The hub gene FAM3b and its prognostic value in TNBC. (A) Number of trees showing the importance proportion of SASP regulator genes. (B) FAM3B expression in paired tumor and normal tissues in BRCA from the TCGA database. (C) FAM3B expression according to the molecular subtypes of breast cancer. (D) UMAP plot of intratumoral immune cells and FAM3B showing the correlation between infiltration of different immune cells and FAM3B expression in the Alex, EMTAB8107, GSE150660 and GSE161529 datasets. (E,F) The impact of FAM3B on OS and DFS in breast cancer tissue microarray using Kaplan-Meier analysis.

Article Snippet: IHC staining of FAM3B protein expression in the tissue microarray was performed by incubation with rabbit polyclonal antibodies against human FAM3B antibody (27131-1-AP, Proteintech, 1:200) overnight, followed by incubation with goat monoclonal antibody against rabbit antibody (111-035-003, JACKSON, 1:1,000) for 1 h at room temperature.

Techniques: Expressing, Microarray

Journal: Human Cell

Article Title: Hsa_circ_0000437 promotes the progression of rheumatic valvular heart disease by activating the mitogen-activated protein kinase signaling pathways after sponging let-7f-5p and targeting RAS-like proto-oncogene B

doi: 10.1007/s13577-025-01331-7

Figure Lengend Snippet: hsa_circ_0000437/let-7f-5p/RALB axis promotes inflammatory transformation during RVHD progression by activating the MAPK signaling pathway. Western blotting assay of MAPK14, ERK, JNK, and RALB proteins in hVICs cells transfected with si-hsa_circ_0000437 ( A ), pcDNA3.1-hsa_circ_0000437 ( B ), let-7f-5p mimic and pcDNA3.1-hsa_circ_0000437 ( C ), si-RALB, pcDNA3.1-hsa_circ_0000437 and let-7f-5p inhibitor ( D ). Numbers indicate the relative expression of proteins, and GAPDH serves as an internal reference. E Diagram of the regulatory mechanism of hsa_circ_0000437/let-7f-5p/RALB axis promoting the occurrence and development of RVHD, the target hsa_circ_0000437 was identified by microarray and confirmed in the cytoplasm that RALB could promote inflammatory transformation during RVHD by activating MAPK signaling pathway. RALB was also inhibited by let-7f-5p and enhanced by ceRNA hsa_circ_0000437 (some elements were created using BioRender.com). n = 3, n stands for biological replicates

Article Snippet: MAPK14 (Cat No. 14064–1-AP), extracellular regulated protein kinases (ERK) (Cat No. 11257–1-AP), c-Jun N-terminal kinase (JNK) (Cat No. 51153–1-AP), RAS-like proto-oncogene B (RALB) (Cat No. 12340–1-AP), and goat anti-rabbit IgG and goat anti-mouse IgG secondary antibody were purchased from a subsidiary of Proteintech Wuhan Sanying Biotechnology Co., LTD (China).

Techniques: Transformation Assay, Western Blot, Transfection, Expressing, Microarray

Journal: Journal of Leukocyte Biology

Article Title: The generation of macrophages with anti-inflammatory activity in the absence of STAT6 signaling

doi: 10.1189/jlb.2A1114-560R

Figure Lengend Snippet: (A) Chemokine and cytokine secretion by BMDMs was measured by a proteome profiler membrane antibody array. Supernatants from nonstimulated macrophages were compared with macrophages treated with 10 ng/ml LPS or a combination of LPS and 100 µl Ova-IC (L+I), 200 nM PGE2 (L+P), 200 μM Ado (L+A), or 20 ng/ml IL-4 for 12 h. The proteins that are of interest to this study are indicated in circles, and the letters and numbers are provided to identify the position of the analyte in the membrane. The profiling was done on pooled supernatants collected from 3 separate experiments from 3 different mice. (B). Mean fold differences in intensity of the duplicate samples for relevant analytes are compared with supernatants from nonstimulated. The alphanumeric values within parentheses indicate their position in the membrane array. BCA-1, B Cell-attracting chemokine-1; TCA-3, T cell activation-3; KC, keratinocyte-derived chemokine.

Article Snippet: Membrane protein array Mouse cytokine antibody array membranes [Proteome Profiler Antibody Array (R&D Systems] were used to assess the relative differences of 40 different cytokines and chemokines in cell culture supernatants.

Techniques: Membrane, Ab Array, Activation Assay, Derivative Assay

Journal: Journal of integrative neuroscience

Article Title: Involvement of CXCL10 in Neuronal Damage under the Condition of Spinal Cord Injury and the Potential Therapeutic Effect of Nrg1.

doi: 10.31083/j.jin2204096

Figure Lengend Snippet: Fig. 1. Expression of Cxcl10 and Nrg1 genes in microarray datasets GSE42828 and GSE93561. (A) Gene expression matrices were

Article Snippet: Following incubation with antibody dilution buffer (Cat no. A1800, Solarbio, Beijing, China), sections were treated with a mixture of the primary rabbit anti-mouse CXCL10 antibody (10937-1-AP, ProteinTech, Rosemont, IL, USA; 1:200) and mouse anti-mouse βIII-Tubulin antibody (sc-80005; Santa Cruz, Dallas, TX, USA; 1:200) [22].

Techniques: Expressing, Microarray, Gene Expression

Journal: Journal of integrative neuroscience

Article Title: Involvement of CXCL10 in Neuronal Damage under the Condition of Spinal Cord Injury and the Potential Therapeutic Effect of Nrg1.

doi: 10.31083/j.jin2204096

Figure Lengend Snippet: Fig. 3. Effect of CXCL10 and Nrg1 on HT22 cells or NSC34 cells viability. (A–C) Effect of CXCL10 (A) and Nrg1 (B), as well as

Article Snippet: Following incubation with antibody dilution buffer (Cat no. A1800, Solarbio, Beijing, China), sections were treated with a mixture of the primary rabbit anti-mouse CXCL10 antibody (10937-1-AP, ProteinTech, Rosemont, IL, USA; 1:200) and mouse anti-mouse βIII-Tubulin antibody (sc-80005; Santa Cruz, Dallas, TX, USA; 1:200) [22].

Techniques:

Journal: Journal of integrative neuroscience

Article Title: Involvement of CXCL10 in Neuronal Damage under the Condition of Spinal Cord Injury and the Potential Therapeutic Effect of Nrg1.

doi: 10.31083/j.jin2204096

Figure Lengend Snippet: Fig. 4. Effect of CXCL10 and Nrg1 on HT22 cells and NSC34 cells after scratch injury. (A,C) Effect of CXCL10 and Nrg1 on HT22 cells after scratch injury. n = 4 per group. Scar bar: 200 µm. S.E.M. (***, Nrg1 + CXCL10 group vs. control group, p < 0.001;

Article Snippet: Following incubation with antibody dilution buffer (Cat no. A1800, Solarbio, Beijing, China), sections were treated with a mixture of the primary rabbit anti-mouse CXCL10 antibody (10937-1-AP, ProteinTech, Rosemont, IL, USA; 1:200) and mouse anti-mouse βIII-Tubulin antibody (sc-80005; Santa Cruz, Dallas, TX, USA; 1:200) [22].

Techniques: Control

Journal: Journal of integrative neuroscience

Article Title: Involvement of CXCL10 in Neuronal Damage under the Condition of Spinal Cord Injury and the Potential Therapeutic Effect of Nrg1.

doi: 10.31083/j.jin2204096

Figure Lengend Snippet: Fig. 5. Effect of CXCL10 and Nrg1 on the molecular expression of HT22 cells. (A) Representative images of pErbB4, pERK1/2, cleaved caspase 9, and cleaved caspase 3. (B–E) Quantitative results of pErbB4, pERK1/2, cleaved caspase 9, and cleaved caspase 3. n

Article Snippet: Following incubation with antibody dilution buffer (Cat no. A1800, Solarbio, Beijing, China), sections were treated with a mixture of the primary rabbit anti-mouse CXCL10 antibody (10937-1-AP, ProteinTech, Rosemont, IL, USA; 1:200) and mouse anti-mouse βIII-Tubulin antibody (sc-80005; Santa Cruz, Dallas, TX, USA; 1:200) [22].

Techniques: Expressing

Journal: Journal of integrative neuroscience

Article Title: Involvement of CXCL10 in Neuronal Damage under the Condition of Spinal Cord Injury and the Potential Therapeutic Effect of Nrg1.

doi: 10.31083/j.jin2204096

Figure Lengend Snippet: Fig. 6. Effect of CXCL10 and Nrg1 on the molecular expression of NSC34 cells. (A) Representative images of pErbB4, pERK1/2, cleaved caspase 9, and cleaved caspase 3. (B–E) Quantitative results of pErbB4, pERK1/2, cleaved caspase 9, and cleaved caspase 3. n

Article Snippet: Following incubation with antibody dilution buffer (Cat no. A1800, Solarbio, Beijing, China), sections were treated with a mixture of the primary rabbit anti-mouse CXCL10 antibody (10937-1-AP, ProteinTech, Rosemont, IL, USA; 1:200) and mouse anti-mouse βIII-Tubulin antibody (sc-80005; Santa Cruz, Dallas, TX, USA; 1:200) [22].

Techniques: Expressing

Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet: FGFR4 is highly expressed in RMS and other cancers, with low expression in healthy tissue (A) High expression of FGFR4 mRNA is found in both FP-RMS and FN-RMS tumors and cell lines compared with other pediatric cancers and healthy tissues. Expression levels measured as FPKM (fragments per kilobase of transcript per million mapped reads) for FGFR4 are summarized in violin plots with medians and quartiles. ASPS, alveolar soft part sarcoma; CCSK, clear cell sarcoma of the kidney; DSRCT, desmoplastic small round cell tumor; EWS, Ewing sarcoma; HBL, hepatoblastoma; ML, melanoma; NB, neuroblastoma; OS, osteosarcoma; SS, synovial sarcoma; UDS, undifferentiated sarcoma; WT, Wilms tumor; YST, yolk sac tumor). (B) FGFR4 mRNA expression in TCGA data shows highest expression in liver hepatocellular carcinoma (LIHC), cholangiocarcinoma (CHOL), and individual tumors of other types. Abbreviations are as per TCGA ( https://gdc.cancer.gov/resources-tcga-users/tcga-code-tables/tcga-study-abbreviations ). (C) Representative images of immunohistochemistry (IHC) for FGFR4 show high expression in RMS, with minimum or no expression in healthy organs. H score displayed in bottom right corner, and scale bars, 200 μm. (D) Summary of membrane-staining H score of FGFR4 IHC of RMS and healthy tissues. Values represent mean ± SEM (error bars). (E) Representative flow cytometry plots show differential levels of FGFR4 expression on FP-RMS or FN-RMS cell lines. Mean fluorescence intensity (MFI) of FGFR4 on indicated RMS cells is listed in the right table, stained with phycoerythrin (PE)-conjugated anti-human FGFR4 antibody or mouse IgG1 isotype control.

Article Snippet: FGFR4 (D3B12) XP® Rabbit mAb , Cell Signaling Technology , Cat# CST8562; RRID:AB_10891199.

Techniques: Expressing, Wilms Tumor Assay, Immunohistochemistry, Membrane, Staining, Flow Cytometry, Fluorescence, Control

Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet: PAX3-FOXO1 establishes a super-enhancer at the FGFR4 locus, and RMSs are dependent on FGFR4 for survival (A) PAX3-FOXO1 (top) and H3K27ac (bottom) ChIP-seq at the FGFR4 locus in FP-RMS cell lines and tumors (orange), FN-RMS cell lines and tumors (blue), and human skeletal muscle cell lines and tissues (gold). (B) Top: FGFR4 expression is induced in fibroblasts after introduction of PAX3-FOXO1. Bottom: ChIP-seq demonstrates that PAX3-FOXO1 protein opens chromatin and establishes a super-enhancer at the FGFR4 locus. Open chromatin was assayed by DNase hypersensitivity; binding of PAX3-FOXO1 and BRD4 as well as chromatin H3K27ac status were assayed by ChIP-seq in human fibroblasts with or without PAX3-FOXO1. (C) Average dependency score of RMS for FGFR4 was found to be the lowest among all human cancers, suggesting the highest dependency of RMS on this receptor for survival.

Article Snippet: FGFR4 (D3B12) XP® Rabbit mAb , Cell Signaling Technology , Cat# CST8562; RRID:AB_10891199.

Techniques: ChIP-sequencing, Expressing, Binding Assay

Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet: Development and characterization of a specific FGFR4 binder from the monoclonal murine antibody 3A11 (A) Structure of the mouse anti-FGFR4 mouse monoclonal antibody (mAb) 3A11. (B) Representative flow cytometry plots show the specificity of 3A11 mAb by staining with FGFR4 + cell lines, FGFR4 KO RH30, or FGFR4 − cell 7250. Mouse IgG (msIgG) is used as isotype control for this mouse antibody 3A11. (C) Structure of anti-FGFR4 antibody 3A11 in scFvFc format fused to the human IgG1 Fc region. (D) Representative flow cytometry plots show 3A11 scFvFc chimeric antibody specifically binds to FGFR4 + cell lines RH30, RH4, and RMS559 but not to the RH30 FGFR4-KO, the RH4 FGFR4-KO, or fibroblast 7250 lines. Human IgG (huIgG) as an isotype control for 3A11 scFvFc. MFI fold change shown as orange font calculated by formula [ M F I ( s c F v F c ) − M F I ( h u I g G i s o t y p e ) ] / M F I ( h u I g G i s o t y p e ) . (E) Binding avidity of FGFR4 scFvFc, using 2-to-1 binding model and global fitting analysis, demonstrates the dissociation constant (K D ) of 3A11 scFvFc against FGFR4 ECD is 4.17 nM. (F) ELISA shows 3A11 scFvFc only recognizes human FGFR4 but not human FGFR1–3 or mouse FGFR4. (G) Flow cytometry using 3A11 scFvFc shows FGFR4 is expressed in several RMS cell lines at various levels with higher expression in FP-RMS compared with in FN-RMS. MFI of 3A11 scFvFc or isotype control huIgG staining on above cells is shown in the table on the right.

Article Snippet: FGFR4 (D3B12) XP® Rabbit mAb , Cell Signaling Technology , Cat# CST8562; RRID:AB_10891199.

Techniques: Flow Cytometry, Staining, Control, Binding Assay, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet: Clinical-grade 3A11 CAR T cells show specific cytotoxicity to FGFR4 + cells (A) Schematic of 3A11 CAR construct targeting FGFR4. HTM, hinge and transmembrane domain; hu tEGFR, human truncated EGFR. (B) Cytotoxicity assays of 3A11 CAR T cells show potent killing activity toward target RMS cells at an E:T ratio of 0.75:1 in a xCELLigence Real-Time Cell Analysis (RTCA). Vertical black arrows show the time point for adding CAR T cells into a plate seeded with target cells. Representative of n = 3 independent experiments with n = 3 individual donors for (B)–(D). Values represent mean ± SD (standard deviation, error bars). (C) Cytotoxic assay shows 3A11 CAR does not cause cytolysis to FGFR4-KO or FGFR4 − cells at an E:T ratio of 0.75:1. Values represent mean ± SD (error bars). (D) Cytokine release assay shows 3A11 CAR T cells only release high level of IFN-γ when cocultured with FGFR4-expressing RMS cells (RH30, RH4, or RMS559) rather than the respective FGFR4-KO cell lines or 7250. Values represent mean ± SEM (error bars). Two-way ANOVA is used to compare secreted IFN-γ by mock T or 3A11 CAR T cells cocultured with the target cells by calculating the p value. ∗p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001; ns: no significant difference.

Article Snippet: FGFR4 (D3B12) XP® Rabbit mAb , Cell Signaling Technology , Cat# CST8562; RRID:AB_10891199.

Techniques: Construct, Activity Assay, Cell Analysis, Standard Deviation, Release Assay, Expressing

Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet: Low or absence of cell surface FGFR4 in primary human cells does not induce cytokine release when cocultured with 3A11 CAR T cells (A) Representative flow cytometry plots show low or absence of FGFR4 cell surface expression on human cardiomyocytes, renal epithelial cells, renal cortical epithelial cells, renal proximal epithelial cells, HEK293, cholangiocytes, and hepatocytes pooled from 10 individuals using 3A11 scFvFc. MFI of 3A11 scFvFc or huIgG1 isotype control staining on these primary cells is shown in the top right table. Relative FGFR4 expression levels on these primary cells compared with RH30 cells are calculated as R e l a t i v e e x p r e s s i o n = [ M F I ( s c F v F c ) − M F I ( h u I g G i s o t y p e ) ] P r i m a r y c e l l [ M F I ( s c F v F c ) − M F I ( h u I g G i s o t y p e ) ] R H 30 . (B) Log 2 ratio of cytokine (IFN-γ, IL-2, and TNF-α) release in the supernatant, by 3A11 CAR T cells cocultured with primary cells, as indicated, in their respective media, compared with the RMS cell RH30. Cell line 7250 serves as a FGFR4 − control. Values represent n = 3 independent experiments with n = 3 individual donors. Values represent mean ± SEM (error bars).

Article Snippet: FGFR4 (D3B12) XP® Rabbit mAb , Cell Signaling Technology , Cat# CST8562; RRID:AB_10891199.

Techniques: Flow Cytometry, Expressing, Control, Staining

Journal: Cell Reports Medicine

Article Title: Preclinical development of a chimeric antigen receptor T cell therapy targeting FGFR4 in rhabdomyosarcoma

doi: 10.1016/j.xcrm.2023.101212

Figure Lengend Snippet:

Article Snippet: FGFR4 (D3B12) XP® Rabbit mAb , Cell Signaling Technology , Cat# CST8562; RRID:AB_10891199.

Techniques: Purification, Control, Recombinant, Labeling, Virus, Microarray, Cytokine Assay, Transfection, Chromatin Immunoprecipitation, ChIP-sequencing, Plasmid Preparation, Software, Imaging

Journal: Stem Cell Research & Therapy

Article Title: Oxidant therapy improves adipogenic differentiation of adipose-derived stem cells in human wound healing

doi: 10.1186/s13287-021-02336-3

Figure Lengend Snippet: Characterization of the macrophage activation model. a THP1 cell treatment protocol for in vitro polarization of macrophages. b Morphology (phase contrast, scale bar 100μm) of THP1-derived macrophages on day of CM harvest. c Heatmap of log2 cytokine protein levels in differentially activated macrophages. The dendrogram represents the result from a complete linkage hierarchical clustering of mean-centered expression levels based on Euclidian similarity. d mRNA-expression levels of macrophage-selective markers (M (IFNG/LPS) : CD80, IL1B, and M (IL4/IL13) : CD200R, TGM2). Data were normalized to the 18S rRNA reference gene and shown as mean –ΔCt. Asterisks indicate p values < 0.05 (*), p <0.01 (**), and p <0.001 (***). Statistical significance ( n =5) was determined by using one-way ANOVA and Dunnett’s test for multiple comparison

Article Snippet: Macrophage-CM was loaded onto RayBio® Human Cytokine G5 Antibody Microarray Glass Chip (RayBiotech, Norcross, GA) which facilitates the detection of 80 targets.

Techniques: Activation Assay, In Vitro, Derivative Assay, Expressing

Journal: Mediators of Inflammation

Article Title: LR12 Promotes Liver Repair by Improving the Resolution of Inflammation and Liver Regeneration in Mice with Thioacetamide- (TAA-) Induced Acute Liver Failure

doi: 10.1155/2021/2327721

Figure Lengend Snippet: LR12 promoted hepatocyte regeneration via CCL20 secreted by macrophages. (a) The cytokine protein microarray of the supernatant from macrophages. (b) ELISA of CCL20 in the supernatant from macrophages. (c) The mRNA level of CCL20 in macrophages. (d) CLSM showing F4/80 and CCL20 staining in the liver tissues. (e) Western blot analysis of PCNA in LO2 cells stimulated with CCL20. Data were presented as the mean ± standard deviation (SD). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 versus control group.

Article Snippet: The RayBio Human Cytokine Antibody Array Kit (QAH-TH17-1-1 microarray; RayBiotech, Norcross, GA, USA) was utilized to identify the cytokines based on instructions provided by the manufacturer.

Techniques: Microarray, Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Standard Deviation