Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 reduces Sema4D levels to elevate cortical strength in mice with type 2 diabetes mellitus (T2DM). (A) Schedule of enzyme-linked immunosorbent assay (ELISA) and the corresponding groups. (B) ELISA to determine the levels of ALP, Runx-2, OCN, OPN, and Sema4D in control mice, mice with T2DM, mice with T2DM treated with metformin (Met)/exendin-4 (Ex-4), and GLP-1R knockdown T2DM mice treated with Ex-4. n=6 mice per group. ALP: Alkaline Phosphatase; Runx-2: Runt-related transcription factor 2; OCN: Osteocalcin; OPN: Osteopontin; Sema4D: Semaphorin 4D. (C) Schedule of micro-CT to determine changes in trabecular bone. (D) Micro-CT images representing trabecular formation in control mice, mice with T2DM, mice with T2DM treated with Ex-4, mice with T2DM treated with anti-Sema4D, and mice with T2DM treated with a combination of Ex-4 and anti-Sema4D within a region of interest (ROI). n=6 mice per group. (E) Quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 mice per group. (F, G) Micro-CT images representing cortical bone formation in control mice, mice with T2DM, mice with T2DM treated with Ex-4, mice with T2DM treated with anti-Sema4D, and mice with T2DM treated with a combination of Ex-4 and anti-Sema4D within an ROI, and quantitative analysis of micro-CT images. n=6 mice per group. (H) Effects of exendin-4 or anti-Sema4D on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 mice per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the regulatory role of Sema4D in cancellous bone loss and impaired bone defect regeneration in mice, mice were randomly divided into 5 groups (n = 6/group): control group, Sema4D group (10 ng/mL), Sema4D group (15 ng/mL), Sema4D (10 ng/mL) + metformin (300 mg/kg/day, MCE) group and Sema4D (15 ng/mL) + metformin (300 mg/kg/day, MCE) group.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Knockdown, Micro-CT, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: The combination of metformin and exendin-4 can neutralize the effect of Sema4D. (A) Schematic chart of the sample preparation of micro-CT and 3-point bending experiments to determine the effect of the combination of metformin and exendin-4. (B) Micro-CT images representing trabecular formation in control mice and in mice treated with metformin, exendin-4, the combination of metformin and exendin-4, and anti-Sema4D within a region of interest (ROI), and quantitative analysis of micro-CT images. BV: bone volume; TV: tissue volume; Tb.N: trabecular number; Tb.Th: trabecular thickness; Tb.Sp: trabecular spacing; BS/BV: bone superficial area/bone volume. n=6 per group. (C) Micro-CT images representing cortical formation in control mice and in mice treated with metformin, exendin-4, the combination of metformin and exendin-4, and anti-Sema4D within an ROI, and quantitative analysis of micro-CT images. n=6 per group. (D) Effects of metformin, exendin-4, the combination of metformin and exendin-4, or anti-Sema4D on the biomechanical properties of femoral diaphysis evaluated using the 3-point bending test. Femoral maximum load (N), stiffness (N/mm), maximum stress (MPa), and Young’s modulus (GPa). n=6 per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the regulatory role of Sema4D in cancellous bone loss and impaired bone defect regeneration in mice, mice were randomly divided into 5 groups (n = 6/group): control group, Sema4D group (10 ng/mL), Sema4D group (15 ng/mL), Sema4D (10 ng/mL) + metformin (300 mg/kg/day, MCE) group and Sema4D (15 ng/mL) + metformin (300 mg/kg/day, MCE) group.

Techniques: Sample Prep, Micro-CT, Control, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 promotes pseudopodia numbers, leading to the stretching and wider spread of BMSCs via CRMP2. (A) Cell morphology observed using fluorescence microscopy. ImageJ was used to split color channels to observe the pseudopodium in human bone mesenchymal stem cells (hBMSCs). n=5 per group. White arrows indicate the pseudopodium in BMSCs. (B) Immunofluorescence staining to determine F-actin expression in BMSCs. n=5 per group. (C) Western blotting to determine F-actin expression in different groups. n=3 per group. (D) Proteomics analysis to determine the expression of different phosphorylated proteins. (E) Cell morphology observed using fluorescence microscopy. ImageJ was used to split color channels to observe the pseudopodium in BMSCs. n=5 per group. White arrows indicate the pseudopodium in BMSCs. (F) Immunofluorescence staining showing F-actin in different groups. n=5 per group. (G) Immunofluorescence staining indicating CRMP2 expression in BMSCs. n=5 per group. CRMP2: Collapsin Response Mediator Protein 2. (H) Western blotting to determine ALP, Runx-2, and Osterix expression in BMSCs. n=3 per group. (I) Alizarin red staining to analyze mineral deposits in BMSCs with DPYSL2 KD/OE. n=3 per group. DPYSL2 KD/OE: Dihydropyrimidinase like protein 2 knockdown/overexpression. (J) Western blotting to determine pCRMP2 expression in BMSCs with Sema4D or exendin-4. n=3 per group. (K) Immunofluorescence staining to determine pCRMP2 expression in BMSCs. n=5 per group. **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the regulatory role of Sema4D in cancellous bone loss and impaired bone defect regeneration in mice, mice were randomly divided into 5 groups (n = 6/group): control group, Sema4D group (10 ng/mL), Sema4D group (15 ng/mL), Sema4D (10 ng/mL) + metformin (300 mg/kg/day, MCE) group and Sema4D (15 ng/mL) + metformin (300 mg/kg/day, MCE) group.

Techniques: Fluorescence, Microscopy, Immunofluorescence, Staining, Expressing, Western Blot, Knockdown, Over Expression, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Exendin-4 activates bone formation via the GLP-1R/PI3K/GSK-3β/CRMP2 signaling pathway. (A) Western blotting results of pCRMP2 and F-actin expression in BMSCs with GSK-3β knockdown or overexpression. n=3 per group. GSK-3β: Glycogen Synthase Kinase 3 beta. (B) Immunofluorescence staining of F-actin and pCRMP2 expression in BMSCs. n=5 per group. (C) Western blotting to determine pCRMP2 expression after Sema4D treatment with or without GSK-3β knockdown. n=3 per group. (D) Western blotting to determine pCRMP2 expression after exendin-4 treatment with or without GSK-3β knockdown. n=3 per group. (E) Western blotting to determine the expression of pGSK-3β, pCRMP2, and F-actin after Akt knockdown or overexpression. n=3 per group. Akt: Protein Kinase B/ Ak strain transforming. (F) Western blotting to determine the expression of pGSK-3β, pCRMP2, and F-actin after PI3K knockdown or overexpression. n=3 per group. PI3K: Phosphoinositide 3-Kinase. (G) Western blotting to determine the expression of pGSK-3β after exendin-4 treatment with or without PI3K knockdown and overexpression. n=3 per group. (H) Western blotting to determine P-PI3K expression after exendin-4 treatment with or without GSK-3β knockdown and overexpression. n=3 per group. (I) Western blotting to determine pGSK-3β expression after Sema4D treatment with or without PI3K knockdown and overexpression. n=3 per group. (J) Western blotting to determine pPI3K expression after Sema4D treatment with or without GSK–3β knockdown or overexpression. n=3 per group. *p < 0.05; **p < 0.01; ***p < 0.001. Data are presented as mean ± standard deviation.

Article Snippet: To investigate the regulatory role of Sema4D in cancellous bone loss and impaired bone defect regeneration in mice, mice were randomly divided into 5 groups (n = 6/group): control group, Sema4D group (10 ng/mL), Sema4D group (15 ng/mL), Sema4D (10 ng/mL) + metformin (300 mg/kg/day, MCE) group and Sema4D (15 ng/mL) + metformin (300 mg/kg/day, MCE) group.

Techniques: Western Blot, Expressing, Knockdown, Over Expression, Immunofluorescence, Staining, Standard Deviation

Journal: Journal of Advanced Research

Article Title: Discovery and identification of semaphorin 4D as a bioindicator of high fracture incidence in type 2 diabetic mice with glucose control

doi: 10.1016/j.jare.2025.03.014

Figure Lengend Snippet: Metformin modulates the expression of plexin B1 and GLP-1R in hBMSCs. (A) Differential expression of miRNAs among hBMSCs, hBMSCs+Sema4D, and hBMSCs+Sema4D+Met. MiR-140-3p and miR-3657 showing a significant increase and reduced expression in hBMSCs treated with Met. (B) Differential expression of miR-140-3p, plxnb1, and GLP-1R in different groups. n=3 per group. (C, D) Western blotting for Plexin B1 and GLP-1R expression in BMSCs, BMSCs+Sema4D, and BMSCs+Sema4D+Met. n=3 per group. (E) MiR-3657 expression in BMSCs, BMSCs+Sema4D, and BMSCs+Sema4D+Met. n=3 per group. (F, G) GLP-1R and Plexin B1 levels in BMSCs (con, miR-140-mimics/inhibitor, miR-3657-mimics/inhibitor). n=3 per group. (H) GLP-1R levels in BMSCs (con, miR-140-mimics, miR-140-mimics + miR-3657 inhibitor, miR-140-3p+miR-3657-mimics). n=3 per group.

Article Snippet: To investigate the regulatory role of Sema4D in cancellous bone loss and impaired bone defect regeneration in mice, mice were randomly divided into 5 groups (n = 6/group): control group, Sema4D group (10 ng/mL), Sema4D group (15 ng/mL), Sema4D (10 ng/mL) + metformin (300 mg/kg/day, MCE) group and Sema4D (15 ng/mL) + metformin (300 mg/kg/day, MCE) group.

Techniques: Expressing, Quantitative Proteomics, Western Blot

Journal: Journal of advanced research

Article Title: Sympathetic hyperinnervation drives abdominal aortic aneurysm development by promoting vascular smooth muscle cell phenotypic switching.

doi: 10.1016/j.jare.2024.05.028

Figure Lengend Snippet: Fig. 7. Elevated Sema4D expression in AAA. (A-C) Heatmap illustrating expression profiles of nerve guidance factors in aorta samples from Ang II-infused ApoE-/- mice and saline-infused ApoE-/- mice. (D) Dot plots showing expression of nerve guidance factors in macrophage from angiotensin II–induced AAA tissue (AAA) and normal aorta tissues (Normal). (E-F) Western blots and quantification analysis of Sema4D protein levels in human AAA samples and control samples (n = 6/group) and aortas from Ang II- induced AAA tissues and saline-induced aorta tissues (n = 6/group). (G-H) Representative immunohistochemical staining and corresponding quantification of Sema4D levels in Ang II-induced AAA models and control (n = 5/group; scale bars = 200 and 50 lm). (I-J) Flow cytometry analysis of Sema4D+CD68+ cells dissociated from saline-induced aorta tissues and Ang II-induced AAA tissues (n = 4/group). (K) Representative confocal images of Sema4D (red) colocalized with CD68 (green)–positive macrophages in aorta tissues from Ang II-infused or saline-infused ApoE-/- mice (scale bars = 50 lm). (L) Representative confocal images of Sema4D (red) colocalized with TRAP (green)–positive osteoclast-like cells in aorta tissues from Ang II-infused or saline-infused ApoE-/- mice (scale bars = 50 lm). (M) Secreted protein levels of Sema4D from conditional medium of macrophages treated with vehicle, TNF-a, CaPO4 and TNF-a + CaPO4 (n = 5/group). Parametric paired t test for Fig. 7F(left), parametric unpaired t test for Fig. 7F(right), 7H, 7J and one-way ANOVA with a post Bonferroni’s multiple comparisons test for 7 M. **P <0.01, ***P <0.001, ****P <0.0001.

Article Snippet: Sema4D knockout (Sema4D-/-) mice were purchased from Cyagen, maintained in the animal facilities of Southern Medical University on standard rodent chow, and switched to a Western diet containing 0.2 % cholesterol by weight (TD.88137, Harlan Teklad) after AAV-PCSK9 injection.

Techniques: Expressing, Saline, Western Blot, Control, Immunohistochemical staining, Staining, Flow Cytometry

Journal: Journal of advanced research

Article Title: Sympathetic hyperinnervation drives abdominal aortic aneurysm development by promoting vascular smooth muscle cell phenotypic switching.

doi: 10.1016/j.jare.2024.05.028

Figure Lengend Snippet: Fig. 8. Sema4D deletion represses AAA formation by inhibiting sympathetic hyperinnervation. (A) Representative photographs showing macroscopic features of aorta. (B) The AAA incidence in Wildtype group (n = 21) and Sema4D-/- group (n = 22). (C) The survival curve from indicated group. (D) The measurements of maximum aortic diameter. (E) Representative macroscopic images of aorta sections stained with H&E, VVG and Masson trichrome (scale bars = 200 and 50 lm). (F) Elastin degradation score in aortas (n = 10/group). (G) Percentage of collagen area in aortas (n = 8/group). (H-I) Western blotting and quantification of GAP43 and TH in aortic tissues (n = 6/group). (J-K) Representative immunofluorescence images and analysis of b-Tubulin III in PC12 cells treated with monocyte-conditioned medium or osteoclast-conditioned medium, with or without the addition of functional blocking antibodies (n = 6/group; scale bars = 25 lm). Mono-CM, monocyte-conditioned medium; OC-CM, osteoclast-conditioned medium; ab, antibody. (L-M) Representative immunofluorescence images and analysis of b-Tubulin III in rSema4D-treated PC12 with si-NC or si-Plxnb1 (scale bars = 25 lm; n = 6/group). Parametric unpaired t test for Fig. 8D, 8G, 8I, and 8M, Fisher’s exact test for Fig. 8B, log-rank (Mantel-Cox) test for Fig. 8C, nonparametric Mann-Whitney U test for Fig. 8F, and One-way ANOVA with a post Bonferroni’s multiple comparisons test for Fig. 8K. *P <0.05, **P <0.01, ****P <0.0001.

Article Snippet: Sema4D knockout (Sema4D-/-) mice were purchased from Cyagen, maintained in the animal facilities of Southern Medical University on standard rodent chow, and switched to a Western diet containing 0.2 % cholesterol by weight (TD.88137, Harlan Teklad) after AAV-PCSK9 injection.

Techniques: Staining, Western Blot, Functional Assay, Blocking Assay, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: The involvement of the primo vascular system in local enteritis and its modification by electroacupuncture

doi: 10.3389/fimmu.2022.1072996

Figure Lengend Snippet: Validating the reliability of the sequencing data (n = 3 per group). Representative protein levels of TAP binding protein (TAPBP), toll-like receptor 2 (TLR2), semaphorin 4D (SEMA4D), jagunal homolog 1 (JAGN1), tumor necrosis factor receptor superfamily member 5 (CD40), anti- receptor-type tyrosine-protein phosphatase C (CD45), anti- lysosomal associated membrane protein 1 (LAMP1), HLA (RLA) class II histocompatibility antigen DR alpha chain (HLA-DRA1), and fibrinogen like 1 (FGL1) were detected in the PVS tissues by western blotting. Data were expressed as mean ± SD. Statistical analysis was performed using an unpaired Student’s t-test. * P < 0.05, ** P < 0.01, vs. control group.

Article Snippet: After being incubated in Tris-buffered saline containing 0.1% Tween-20 (TBST) with 5% nonfat milk for 1 h, the membranes were infiltrated with anti-tubulin-α antibody (Proteintech 66031-1-Ig, diluted 1:3000), anti-tumor necrosis factor receptor superfamily member 5 (CD40) antibody (Abclonal A0218, diluted 1:1000), anti-receptor-type tyrosine-protein phosphatase C (CD45) (Huabio ER1901-29, diluted 1:1000), anti-HLA (RLA) class II histocompatibility antigen DR alpha chain (HLA-DRA1) antibody (Abclonal A10863, diluted 1:1000), anti-TAP binding protein (TAPBP) antibody (Abclonal 13357, diluted 1:1000) anti-toll-like receptor 2 (TLR2) antibody (Abclonal A2545, diluted 1:1000), anti-semaphorin 4D (SEMA4D) antibody (Abclonal A10136, diluted 1:1000), anti-fibrinogen like 1 (FGL1) antibody (Abclonal A20335, diluted 1:1000), anti-jagunal homolog 1 (JAGN1) antibody (Santa Cruz sc-515306, diluted 1:1000), and anti-lysosomal associated membrane protein 1 (LAMP1) antibody (CST 9091, diluted 1:1000), respectively, overnight at 4°C.

Techniques: Sequencing, Binding Assay, Membrane, Western Blot, Control

Journal: Immunity

Article Title: Differential IRF8 Transcription Factor Requirement Defines Two Pathways of Dendritic Cell Development in Humans

doi: 10.1016/j.immuni.2020.07.003

Figure Lengend Snippet:

Article Snippet: Human anti-human CD100 APC-Vio770, clone REA316 , Miltenyi Biotec , Cat# 130-104-604; RRID: AB_2654328.

Techniques: Purification, Recombinant, Concentration Assay, Saline, Staining, Antibody Labeling, RNA Sequencing, Software, Sterility

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Semaphorin 4D Contributes to Rheumatoid Arthritis by Inducing Inflammatory Cytokine Production: Pathogenic and Therapeutic Implications

doi: 10.1002/art.39086

Figure Lengend Snippet: Levels of soluble semaphorin 4D (sSema4D) in serum and synovial fluid samples from patients with rheumatoid arthritis (RA), and Sema4D protein expression in RA synovium. A, Soluble Sema4D levels in serum samples from healthy individuals and from 101 patients with RA, 10 patients with osteoarthritis (OA), 10 patients with ankylosing spondylitis (AS), and 34 patients with systemic lupus erythematosus (SLE). B, Levels of sSema4D in synovial fluid samples from 7 RA patients and 10 OA patients. Values in A and B are the mean ± SD. ND = not detectable. * = P < 0.05; ** = P < 0.01; ***= P < 0.001. C, Hematoxylin and eosin (H&E) staining and immunohistochemical staining for Sema4D, CD3, CD20, and CD68 in synovial tissue samples from patients with OA or RA. Images shown are representative of samples from 7 RA patients and 10 OA patients. Original magnification × 40. Ab = antibody.

Article Snippet: Assay ID numbers for TaqMan primer sets (Applied Biosystems) were Hs00174819_m1 for human SEMA45 and HS00192708_m1 for human ADAMTS4 .

Techniques: Expressing, Staining, Immunohistochemical staining

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Semaphorin 4D Contributes to Rheumatoid Arthritis by Inducing Inflammatory Cytokine Production: Pathogenic and Therapeutic Implications

doi: 10.1002/art.39086

Figure Lengend Snippet: Correlations of serum levels of soluble semaphorin 4D (sSema4D) with markers of rheumatoid arthritis disease activity. A, Positive correlation of serum sSema4D levels with the Disease Activity Score in 28 joints (DAS28), the C‐reactive protein (CRP) level, the rheumatoid factor (RF) titer, and the level of urinary deoxypyridinoline (u‐DPD). B, Serum sSema4D levels before and after biologic disease‐modifying antirheumatic drug (DMARD) treatment in 9 good responders and 8 moderate responders or nonresponders according to the European League Against Rheumatism response criteria. *** = P < 0.001. NS = not significant. C, Correlation of change ratio in serum sSema4D levels with change ratio of DAS28 after treatment with biologic DMARDs (n = 17 samples).

Article Snippet: Assay ID numbers for TaqMan primer sets (Applied Biosystems) were Hs00174819_m1 for human SEMA45 and HS00192708_m1 for human ADAMTS4 .

Techniques: Activity Assay

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Semaphorin 4D Contributes to Rheumatoid Arthritis by Inducing Inflammatory Cytokine Production: Pathogenic and Therapeutic Implications

doi: 10.1002/art.39086

Figure Lengend Snippet: Semaphorin 4D (Sema4D) expression, and soluble Sema4D (sSema4D) production with ADAMTS‐4 as the sheddase. A, Histograms of cell surface expression of Sema4D in peripheral blood CD3+, CD19+, and CD14+ cells. Results shown are representative of findings from 5 rheumatoid arthritis (RA) patients and 5 healthy individuals. B, Expression of mRNA for Sema4D in peripheral blood CD3+, CD19+, and CD14+ cells. Results shown are from 5 RA patients and 5 healthy individuals. C, Levels of sSema4D in culture supernatant of THP‐1 cells cultured with recombinant matrix metalloproteinase 3 (MMP‐3), MMP‐9, ADAM‐17, and ADAMTS‐4 (n = 5 samples per group). Results are representative of 3 independent experiments. D, Serum levels of ADAMTS‐4 in 20 RA patients and 16 healthy individuals. E, Synovial fluid levels of ADAMTS‐4 in 7 RA patients and 10 osteoarthritis (OA) patients. Values in B–E are the mean ± SEM. ** = P < 0.01. NS = not significant.

Article Snippet: Assay ID numbers for TaqMan primer sets (Applied Biosystems) were Hs00174819_m1 for human SEMA45 and HS00192708_m1 for human ADAMTS4 .

Techniques: Expressing, Cell Culture, Recombinant

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Semaphorin 4D Contributes to Rheumatoid Arthritis by Inducing Inflammatory Cytokine Production: Pathogenic and Therapeutic Implications

doi: 10.1002/art.39086

Figure Lengend Snippet: Inflammatory cytokine production induced by semaphorin 4D (Sema4D), and elevated ADAMTS‐4 expression by inflammatory cytokines. A, Tumor necrosis factor α (TNFα) and interleukin‐6 (IL‐6) levels in culture supernatant of CD14+ monocytes from rheumatoid arthritis (RA) patients after stimulation with naturally cleaved soluble Sema4D (sSema4D) for 72 hours. Results shown are representative of 3 independent experiments. B, TNFα and IL‐6 levels in culture supernatant of CD14+ monocytes from RA patients after stimulation with naturally cleaved sSema4D for 48 hours with or without anti‐Sema4D antibody. Results shown are representative of 3 independent experiments. C and D, Elevated expression of ADAMTS‐4 mRNA ( C ) and elevated ADAMTS‐4 protein levels ( D ) in primary cultures of TNFα‐ and IL‐6–stimulated synovial cells from RA patients. Data were compiled from 5 independent experiments. Values are the mean ± SEM. * = P < 0.05; ** = P < 0.01. non = not stimulated.

Article Snippet: Assay ID numbers for TaqMan primer sets (Applied Biosystems) were Hs00174819_m1 for human SEMA45 and HS00192708_m1 for human ADAMTS4 .

Techniques: Expressing

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Semaphorin 4D Contributes to Rheumatoid Arthritis by Inducing Inflammatory Cytokine Production: Pathogenic and Therapeutic Implications

doi: 10.1002/art.39086

Figure Lengend Snippet: Blocking of semaphorin 4D (Sema4D) ameliorates severity of collagen‐induced arthritis (CIA) in mice. A, Average arthritis scores of mice with CIA. Anti‐Sema4D or control antibody (Ab) (50 mg/kg) was administered intraperitoneally on days 28 and 35 ( arrows ) (n = 6 mice per group). Data are representative of 3 independent experiments. B, Sections of a mouse ankle joint on day 42 after first immunization. Sections were stained with hematoxylin and eosin (H&E) or Safranin O (SO). Original magnification × 100. C, Average pathologic scores of paw sections on day 42 (n = 6 mice per group). D, Serum levels of tumor necrosis factor α (TNFα) and interleukin‐6 (IL‐6) on day 42 (n = 6 mice per group). Values are the mean ± SEM and are representative of 3 independent experiments. * = P < 0.05; ** = P < 0.01.

Article Snippet: Assay ID numbers for TaqMan primer sets (Applied Biosystems) were Hs00174819_m1 for human SEMA45 and HS00192708_m1 for human ADAMTS4 .

Techniques: Blocking Assay, Control, Staining

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Semaphorin 4D Contributes to Rheumatoid Arthritis by Inducing Inflammatory Cytokine Production: Pathogenic and Therapeutic Implications

doi: 10.1002/art.39086

Figure Lengend Snippet: Correlations between levels of soluble semaphorin 4D and clinical features in the 101 patients with rheumatoid arthritis a

Article Snippet: Assay ID numbers for TaqMan primer sets (Applied Biosystems) were Hs00174819_m1 for human SEMA45 and HS00192708_m1 for human ADAMTS4 .

Techniques: Activity Assay