Journal: The Journal of Immunology Author Choice

Article Title: Citrullination Controls Dendritic Cell Transdifferentiation into Osteoclasts

doi: 10.4049/jimmunol.1800534

Figure Lengend Snippet: Protein citrullination and high PAD activity levels are associated with differentiation plasticity toward OCs in iDCs. (A) PCA indicates clearly distinct protein profiles in DCs obtained from sparse or dense cultures (principal components 1 and 2 are shown). Triplicate samples were analyzed from three independent experiments. (B) Images showing resorption activity on calcium phosphate–coated surface representing OCs that were generated from sparse or dense culture–derived iDCs (original magnification ×40). (C) Higher PAD activity was observed in the lysates of DCs that were obtained from dense cultures. Activity levels are normalized to sparse cultures; the graph represents four individual experiments, each with three replicates. *p < 0.05. (D) Normalized PAD activity levels are shown in cell lysates of iDCs and mature OCs. (E) Immunohistochemistry images indicate 3,3-diaminobenzidene staining of PAD4 and PAD2 enzymes with their respective irrelevant controls during OC development from DC precursors. Slides were counterstained with Mayer’s hemotoxylin and analyzed in light microscopy (original magnification ×100). (F) Citrullinated actin was identified using mass spectrometry in lysates of DCs developing in sparse or dense cultures. The graph shows Mascot score values obtained for the identified peptide; symbols represent three different DC cultures. Mean ± SD values are shown. (G) Reduced DC–OC transdifferentiation in the presence of the PAD inhibitor Cl-amidine.

Article Snippet: Slides were then incubated with rabbit polyclonal anti-PAD2 (Cosmobio, Tokyo, Japan) and anti-PAD4 (Abcam, Cambridge, U.K.) primary Abs, diluted in PBS containing 0.1% saponin for 90 min at room temperature at a final concentration of 0.02 μg/ml for the anti-PAD2 Ab and 1 μg/ml for the anti-PAD4 Ab.

Techniques: Activity Assay, Generated, Derivative Assay, Immunohistochemistry, Staining, Light Microscopy, Mass Spectrometry

Journal: Scientific Reports

Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway

doi: 10.1038/s41598-025-13656-2

Figure Lengend Snippet: Effect of AMF30a on in vitro proliferation of human corneal endothelial cells (hCECs). AMF30a (5µM, PAD2 inhibitor, Cayman, Ann Arbor, Michigan) was applied for 48 h. ( A ) hCECs were identified using immunofluorescence staining of ZO1 and CD166. ( B - C ) Cell area of hCECs were evaluated using cell morphological pictures. Scale bar = 100 μm. ( D and E ) Cell proliferation was measured using CCK-8 cell viability assay and BrdU incorporation assay. ( F ) Cytotoxicity was evaluated using LDH cytotoxicity assay (1.002 ± 0.047 vs. 0.925 ± 0.018). ( G - I ) Cell cycle analysis was assessed using PI staining. (J and K) Wound healing assay was performed using the measurement of scratched ares. Scale bar = 500 μm. ( L and N ) Intracellular oxidative stress levels were measured using DCF-DA assay. Scale bar = 50 μm. * p < 0.05, ** p < 0.01 and **** p < 0.0001.

Article Snippet: The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.

Techniques: In Vitro, Immunofluorescence, Staining, CCK-8 Assay, Viability Assay, BrdU Incorporation Assay, LDH Cytotoxicity Assay, Cell Cycle Assay, Wound Healing Assay

Journal: Scientific Reports

Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway

doi: 10.1038/s41598-025-13656-2

Figure Lengend Snippet: Effect of cytokine on human corneal endothelial cells. ( A ) Immunofluorescence staining of PAD2 shows the distribution of PAD2. Scale bar = 100 μm. ( B and C ) Western blot of PAD2 was used to evaluate the PAD2 levels. ( D and E ) ATF4 levels were evaluated using western blot. ( F ) Immunofluorescence staining of peptidyl-citrulline shows the intensity and distribution of peptidyl-citrulline. Scale bar = 100 μm. * p < 0.05.

Article Snippet: The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.

Techniques: Immunofluorescence, Staining, Western Blot

Journal: Scientific Reports

Article Title: AMF30a promotes survival and function of human corneal endothelial cells by regulating TGF-β/ROCK/HIPPO pathway

doi: 10.1038/s41598-025-13656-2

Figure Lengend Snippet: Schematic diagram summarizing PAD2-mediated signaling in hCECs.

Article Snippet: The cells were treated overnight with either mouse anti-E-cadherin antibody (sc-8426; Santa Cruz Biotechnology, Santa Cruz, CA, USA) or mouse anti-YAP antibody (sc-376830; Santa Cruz Biotechnology), rabbit anti-PAD2 antibody (sc-293271; Santa Cruz Biotechnology), mouse anti-peptidyl-citrulline antibody (MABN328, Merck Millipore, Feltham, UK), and rabbit anti-NF-κB antibody (ab7970, Abcam, 1:100) at 4 °C and then rinsed with PBS.

Techniques:

Journal: Biochimica et Biophysica Acta

Article Title: Modulation of calcium-induced cell death in human neural stem cells by the novel peptidylarginine deiminase–AIF pathway

doi: 10.1016/j.bbamcr.2014.02.018

Figure Lengend Snippet: PAD expression in the developing human central nervous system (brain and spinal cord) and in hNSCs. A) Real time RT-PCR analysis of PAD3 and PAD2 in fetal brains (left panel) and spinal cords (right panel) from human embryos at 42, 63 and 70 days of gestation. PAD2 expression increases while PAD3 expression decreases with development. Human liver was used as a positive control for all PADs. Asterisk indicates statistically significant differences (p < 0.05). B) PAD2 and PAD3 transcript detected by in situ hybridization in human spinal cord at 46 days of gestation (dg). Scale bars are 200 μm. C) PAD2 and PAD3 protein detected by Western blot in developing human brain (Br) and spinal cord (SC); no dramatic change in PAD protein expression is observed.

Article Snippet: The primary antibodies used were rabbit anti-PAD3 (Covalab), rabbit anti-PAD2 (Covalab), rabbit anti-Cleaved Caspase 3 (Cell Signaling), goat anti-AIF (Santa Cruz), mouse anti-ß3-tubulin (Promega), mouse anti-vimentin (Dako), and Alexa 568-conjugated anti-Annexin V (Life Technologies).

Techniques: Expressing, Quantitative RT-PCR, Positive Control, In Situ Hybridization, Western Blot

Journal: Biochimica et Biophysica Acta

Article Title: Modulation of calcium-induced cell death in human neural stem cells by the novel peptidylarginine deiminase–AIF pathway

doi: 10.1016/j.bbamcr.2014.02.018

Figure Lengend Snippet: PADs are expressed in human neural stem cells (hNSCs) and PAD3 inhibition increases hNSC proliferation. A) PAD2 and PAD3 transcript detected by RT-qPCR in hNSC derived from embryonic brain and spinal cord (SC). B) Detection of PAD2 and PAD3 by immunocytochemistry (red) in hNSCs: both proteins are detected in cytoplasm and nucleus (counterstained with Hoechst dye). Scale bars: 25 μm. All pictures are at the same magnification. C) Analysis of cell growth determined by the methylene blue assay after treatment with 100 μM Cl-amidine for 24, 48 or 96 h. Cl-amidine significantly increases hNSC proliferation as compared to controls at 48 and 96 h. * = p < 0.05, ** = p < 0.01 by ANOVA and Student's t -test. D) Analysis of cell growth determined by the methylene blue assay after transfection with siRNA against PAD2 (siPAD2) and PAD3 (siPAD3) or scrambled siRNA. A significant increase in cell growth as compared to controls is observed at 48 h only in cells transfected with siPAD3 (p < 0.05; two-way ANOVA). Error bars indicate SDM; n ≥ 3.

Article Snippet: The primary antibodies used were rabbit anti-PAD3 (Covalab), rabbit anti-PAD2 (Covalab), rabbit anti-Cleaved Caspase 3 (Cell Signaling), goat anti-AIF (Santa Cruz), mouse anti-ß3-tubulin (Promega), mouse anti-vimentin (Dako), and Alexa 568-conjugated anti-Annexin V (Life Technologies).

Techniques: Inhibition, Quantitative RT-PCR, Derivative Assay, Immunocytochemistry, Transfection

Journal: Biochimica et Biophysica Acta

Article Title: Modulation of calcium-induced cell death in human neural stem cells by the novel peptidylarginine deiminase–AIF pathway

doi: 10.1016/j.bbamcr.2014.02.018

Figure Lengend Snippet: Effect of thapsigargin on PAD expression and citrullination in hNSCs. A) RT-qPCR analysis of PAD2 and PAD3 transcripts after thapsigargin treatment: note up-regulation of PAD3, but not PAD2 transcript, in treated cells; * = p < 0.05 by ANOVA and Student's t -test (n ≥ 3; error bars indicate SDM). B) Western blot analysis of citrullinated proteins detected by F95 monoclonal antibody and of citrullinated histone H3 (Cit-H3) following treatment with either Cl-amidine (100 μM) or thapsigargin (5 μM) alone, or both compounds for 24 h. Cl-amidine was added to the culture medium 15 min before thapsigargin treatment. Actin was used as a loading control. Note that PAD activation by thapsigargin results in protein citrullination and this is reduced by the PAD inhibitor, Cl-amidine.

Article Snippet: The primary antibodies used were rabbit anti-PAD3 (Covalab), rabbit anti-PAD2 (Covalab), rabbit anti-Cleaved Caspase 3 (Cell Signaling), goat anti-AIF (Santa Cruz), mouse anti-ß3-tubulin (Promega), mouse anti-vimentin (Dako), and Alexa 568-conjugated anti-Annexin V (Life Technologies).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Activation Assay

Journal: Biochimica et Biophysica Acta

Article Title: Modulation of calcium-induced cell death in human neural stem cells by the novel peptidylarginine deiminase–AIF pathway

doi: 10.1016/j.bbamcr.2014.02.018

Figure Lengend Snippet: PAD3 is the main PAD involved in hNSC cell death. Death/survival of hNSCs treated for 24 h with thapsigargin was determined by the methylene blue assay and staining for the apoptosis marker annexin V. A) Cell survival is significantly (p < 0.05; two-way ANOVA) reduced in hNSCs carrying PAD3-EGFP as compared to cells transfected with PAD3 lacking the active site (ΔPAD3-EGFP), EGFP alone, or no transfection (WT). B) Example of cells expressing PAD3-EGFP (PAD, green) and Annexin-V (AnV, red) counted for quantification (arrows); note the significantly higher percentage of PAD3-EGFP/Annexin-V-positive cells; * = p < 0.05 by ANOVA and Student's t -test. C) Cell survival is significantly (p < 0.05; two-way ANOVA) increased in hNSC transfected with PAD3 but not PAD2 siRNA. Scale bars are 50 μmin B. Error bars indicate SDM; n ≥ 3.

Article Snippet: The primary antibodies used were rabbit anti-PAD3 (Covalab), rabbit anti-PAD2 (Covalab), rabbit anti-Cleaved Caspase 3 (Cell Signaling), goat anti-AIF (Santa Cruz), mouse anti-ß3-tubulin (Promega), mouse anti-vimentin (Dako), and Alexa 568-conjugated anti-Annexin V (Life Technologies).

Techniques: Staining, Marker, Transfection, Expressing

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Reciprocal regulation of protein arginine deiminase 2 and 4 expression in the colonic mucosa of ulcerative colitis

doi: 10.3164/jcbn.23-77

Figure Lengend Snippet: mRNA expression of protein arginine deiminase 2 and 4 in patients with inflammatory bowel diseases. mRNA was isolated from the ileal and colonic mucosa of patients with ulcerative colitis (UC; active disease, n = 20; remitted disease, n = 20) and Crohn’s disease (CD; active disease, n = 10; remitted disease, n = 10). Non-tumorous portions of the colonic mucosa in patients with colon adenoma served as healthy colonic mucosa (HC, n = 4). qRT-PCR analysis of the mRNA expression levels of protein arginine deiminase (PAD)2 and PAD4. Each dot represents the value for each patient. (A) Data are presented as the mean ± SE. ** p <0.01, N.S.; not significant. (B) Correlation between PAD2 and PAD4 mRNA expression in patients with UC and CD. P values and correlation coefficient ( r ) values, as determined by Pearson’s correlation coefficient, are shown.

Article Snippet: PAD2 and PAD4 expression was visualized using the DAKO EnVision+ System (DAKO JAPAN, Tokyo, Japan), as previously described, with rabbit human anti-PAD2 antibody (Proteintech, Rosemont, IL) and anti-PAD4 antibody (GeneTex, Irvine, CA). ( )

Techniques: Expressing, Isolation, Quantitative RT-PCR

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Reciprocal regulation of protein arginine deiminase 2 and 4 expression in the colonic mucosa of ulcerative colitis

doi: 10.3164/jcbn.23-77

Figure Lengend Snippet: mRNA expression of protein arginine deiminase 2 and 4 in patients with different types of ulcerative colitis and Crohn’s disease. mRNA was isolated from the ileal and colonic mucosa of patients with ulcerative colitis (UC; active disease, n = 20; remitted disease, n = 20, A) and Crohn’s disease (CD; active disease, n = 10; remitted disease, n = 10, B). qRT-PCR analysis of the mRNA expression levels of protein arginine deiminase (PAD)2 and PAD4. Each dot represents the value for each patient. Data are presented as the mean ± SE. ** p <0.01.

Article Snippet: PAD2 and PAD4 expression was visualized using the DAKO EnVision+ System (DAKO JAPAN, Tokyo, Japan), as previously described, with rabbit human anti-PAD2 antibody (Proteintech, Rosemont, IL) and anti-PAD4 antibody (GeneTex, Irvine, CA). ( )

Techniques: Expressing, Isolation, Quantitative RT-PCR

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Reciprocal regulation of protein arginine deiminase 2 and 4 expression in the colonic mucosa of ulcerative colitis

doi: 10.3164/jcbn.23-77

Figure Lengend Snippet: Correlation between the mRNA expression of protein arginine deiminase 4 ( PAD4 ) and cytokines in patients with ulcerative colitis (UC). mRNA was isolated from the colonic mucosa of patients with UC. (A) Correlation between the mRNA expression of PAD4 and IFN-α4 , IFN-β , IL12/23p40 , C-X-C motif chemokine ligand 8 ( CXCL8 ), CXCL10 , TNF receptor-associated factor 3 ( TRAF3 ), interferon regulatory factor 3 ( IRF3 ), IRF7 , IL-6 , and TNF -α. (B) Correlation between the mRNA expression of PAD2 and IFN-α4 , IFN-β , CXCL8 , CXCL10 , IL-6 , and TNF -α. Each dot represents the value for each patient. P values and correlation coefficient ( r ) values, as determined by Pearson’s correlation coefficient, are shown.

Article Snippet: PAD2 and PAD4 expression was visualized using the DAKO EnVision+ System (DAKO JAPAN, Tokyo, Japan), as previously described, with rabbit human anti-PAD2 antibody (Proteintech, Rosemont, IL) and anti-PAD4 antibody (GeneTex, Irvine, CA). ( )

Techniques: Expressing, Isolation

Journal: Journal of Clinical Biochemistry and Nutrition

Article Title: Reciprocal regulation of protein arginine deiminase 2 and 4 expression in the colonic mucosa of ulcerative colitis

doi: 10.3164/jcbn.23-77

Figure Lengend Snippet: Correlation between the mRNA expression of protein arginine deiminase 4 ( PAD4 ) and cytokines in patients with Crohn’s disease (CD). mRNA was isolated from the ileal and colonic mucosa of patients with CD. (A) Correlation between the mRNA expression of PAD4 and IFN-α4 , IFN-β , IL12/23p40 , C-X-C motif chemokine ligand 8 ( CXCL8 ), CXCL10 , TNF receptor-associated factor 3 ( TRAF3 ), interferon regulatory factor 3 ( IRF3 ), IRF7 , IL-6 , and TNF-α . (B) Correlation between the mRNA expression of PAD2 and IFN-α4 , IFN-β , CXCL8 , CXCL10 , IL-6 , and TNF-α . Each dot represents the value for each patient. P values and correlation coefficient ( r ) values, as determined by Pearson’s correlation coefficient, are shown.

Article Snippet: PAD2 and PAD4 expression was visualized using the DAKO EnVision+ System (DAKO JAPAN, Tokyo, Japan), as previously described, with rabbit human anti-PAD2 antibody (Proteintech, Rosemont, IL) and anti-PAD4 antibody (GeneTex, Irvine, CA). ( )

Techniques: Expressing, Isolation

Journal: Biology of Reproduction

Article Title: Peptidylarginine deiminase enzymes and citrullinated proteins in female reproductive physiology and associated diseases ^†

doi: 10.1093/biolre/ioac173

Figure Lengend Snippet: PAD2 (green) is expressed in placentas from different species. Placental tissue samples were collected under approval of the Colorado State University IACUC committee (protocol number 1303). Individual sheep, horse, dog, and guinea pig placentas were obtained at gestational days 75 (sheep) and at term (horse, dog, guinea pig) then fixed in 4% paraformaldehyde and paraffin embedded. Five micron sections were processed for H&E staining or routine immunofluorescence using an anti-PAD2 antibody (1:100 ProteinTech, 12110-1-AP) and imaged using Olympus BX63 fluorescence microscope. Blue nuclear staining is DAPI, small inserts are no primary antibody negative controls, and scale bar represents 20 μm. Light blue in zonary and hemochorial is autofluorescence from red blood cells.

Article Snippet: Tumors ( n = 4 /tumor subtype) were examined by immunohistochemistry using an anti-PAD2 antibody (1:100 ProteinTech, 12110-1-AP) or an equal mass of non-specific IgG.

Techniques: Staining, Immunofluorescence, Fluorescence, Microscopy

Journal: Biology of Reproduction

Article Title: Peptidylarginine deiminase enzymes and citrullinated proteins in female reproductive physiology and associated diseases ^†

doi: 10.1093/biolre/ioac173

Figure Lengend Snippet: PAD2 and 4 are expressed in normal human endometrial tissue (n = 2/ out of 4 cores) and a subset of endometrial adenocarcinomas (n = 2/ out of 14 cores). Serial uterine tumor array slides were purchased from US Biomax (T094) and contain 12 cases with 24 cores and include associated pathology grade. Tumor array slides were examined by immunohistochemistry using an anti-PAD2 antibody (1:100 ProteinTech, 12110-1-AP), anti-PAD4 (1:100 Millipore Sigma, P4874) or an equal mass of non-specific IgG (data not shown). Images were taken with a Zeiss AxioVert.A1 with AxioCam MRc and scale bar represents 100 μm.

Article Snippet: Tumors ( n = 4 /tumor subtype) were examined by immunohistochemistry using an anti-PAD2 antibody (1:100 ProteinTech, 12110-1-AP) or an equal mass of non-specific IgG.

Techniques: Immunohistochemistry

Journal: Biology of Reproduction

Article Title: Peptidylarginine deiminase enzymes and citrullinated proteins in female reproductive physiology and associated diseases ^†

doi: 10.1093/biolre/ioac173

Figure Lengend Snippet: PAD2 expression is present in multiple subtypes of pituitary tumors. Pituitary tumor samples including subtype pathology were obtained from Institut D’Investigacions Biomediques August Pi i Sunyer Biobanc (Barcelona, Spain). Samples were de-identified and thus were exempt from University of Wyoming institutional review board (IRB) approval (protocol number 20140814BC00496). Tumors (n = 4 /tumor subtype) were examined by immunohistochemistry using an anti-PAD2 antibody (1:100 ProteinTech, 12110-1-AP) or an equal mass of non-specific IgG. Images were taken with a Zeiss AxioVert.A1 with AxioCam MRc and scale bar represents 50 or 100 μm.

Article Snippet: Tumors ( n = 4 /tumor subtype) were examined by immunohistochemistry using an anti-PAD2 antibody (1:100 ProteinTech, 12110-1-AP) or an equal mass of non-specific IgG.

Techniques: Expressing, Immunohistochemistry

Journal: Arthritis Research & Therapy

Article Title: Rheumatoid arthritis antigens homocitrulline and citrulline are generated by local myeloperoxidase and peptidyl arginine deiminases 2, 3 and 4 in rheumatoid nodule and synovial tissue

doi: 10.1186/s13075-016-1140-9

Figure Lengend Snippet: OA synovial tissue histology. a H&E, ( b ) citrulline and homocitrulline (F95), ( c ) PAD2, ( d ) PAD3, ( e ) PAD4 and ( f ) MPO staining. The scale bar indicates 50 μm. L lumen of synovial cavity, blue arrow synovial lining layer, double-headed arrow synovial stroma

Article Snippet: PAD2 enzyme was stained with anti-human PAD2 antibody 1:250 (ab16478, Abcam) and PAD3 with 1:50 (ab183209, Abcam) after heat-mediated antigen retrieval in Tris-EDTA, pH 9 and myeloperoxidase without antigen retrieval with anti-human myeloperoxidase antibody 1:5000 (A0398, Dako).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: Rheumatoid arthritis antigens homocitrulline and citrulline are generated by local myeloperoxidase and peptidyl arginine deiminases 2, 3 and 4 in rheumatoid nodule and synovial tissue

doi: 10.1186/s13075-016-1140-9

Figure Lengend Snippet: RA synovial tissue histology. a H&E, ( b ) citrulline and homocitrulline (F95), ( c ) PAD2, ( d ) PAD3, ( e ) PAD4 and ( f ) MPO staining. The scale bar indicates 50 μm. L lumen of synovial cavity, blue arrow synovial lining layer, double-headed arrow synovial stroma, black arrow intact neutrophil, arrowhead disrupted neutrophil

Article Snippet: PAD2 enzyme was stained with anti-human PAD2 antibody 1:250 (ab16478, Abcam) and PAD3 with 1:50 (ab183209, Abcam) after heat-mediated antigen retrieval in Tris-EDTA, pH 9 and myeloperoxidase without antigen retrieval with anti-human myeloperoxidase antibody 1:5000 (A0398, Dako).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: Rheumatoid arthritis antigens homocitrulline and citrulline are generated by local myeloperoxidase and peptidyl arginine deiminases 2, 3 and 4 in rheumatoid nodule and synovial tissue

doi: 10.1186/s13075-016-1140-9

Figure Lengend Snippet: Histology of necrotic RA synovial tissue consisting of fibrinoid necrosis and sporadic intact cells. a H&E, ( b ) citrulline and homocitrulline (F95), ( c ) PAD2, ( d ) and PAD3, ( e ) PAD4 and ( f ) MPO staining. The scale bar indicates 50 μm. Arrowhead intact cells, purple double-headed arrow fibrinoid necrosis

Article Snippet: PAD2 enzyme was stained with anti-human PAD2 antibody 1:250 (ab16478, Abcam) and PAD3 with 1:50 (ab183209, Abcam) after heat-mediated antigen retrieval in Tris-EDTA, pH 9 and myeloperoxidase without antigen retrieval with anti-human myeloperoxidase antibody 1:5000 (A0398, Dako).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: Rheumatoid arthritis antigens homocitrulline and citrulline are generated by local myeloperoxidase and peptidyl arginine deiminases 2, 3 and 4 in rheumatoid nodule and synovial tissue

doi: 10.1186/s13075-016-1140-9

Figure Lengend Snippet: Histology of rheumatoid nodule. a H&E, ( b ) citrulline and homocitrulline (F95), ( c ) PAD2, ( d ) PAD3, ( e ) PAD4 and ( f ) MPO staining. The scale bar indicates 50 μm. Purple double-headed arrow fibrinoid necrosis, blue double-headed arrow palisading histiocytes, black double-headed arrow stromal cells

Article Snippet: PAD2 enzyme was stained with anti-human PAD2 antibody 1:250 (ab16478, Abcam) and PAD3 with 1:50 (ab183209, Abcam) after heat-mediated antigen retrieval in Tris-EDTA, pH 9 and myeloperoxidase without antigen retrieval with anti-human myeloperoxidase antibody 1:5000 (A0398, Dako).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: Rheumatoid arthritis antigens homocitrulline and citrulline are generated by local myeloperoxidase and peptidyl arginine deiminases 2, 3 and 4 in rheumatoid nodule and synovial tissue

doi: 10.1186/s13075-016-1140-9

Figure Lengend Snippet: Cellular localization of ( a ) F95 in RA, ( b ) PAD2 in RA, ( c ) PAD3 in RA, ( d ) MPO in RA, ( e ) PAD4 in RA and ( f ) PAD4 staining in OA. The scale bar indicates 50 μm. L lumen of synovial cavity, black arrow intact neutrophil, arrowhead disrupted neutrophil

Article Snippet: PAD2 enzyme was stained with anti-human PAD2 antibody 1:250 (ab16478, Abcam) and PAD3 with 1:50 (ab183209, Abcam) after heat-mediated antigen retrieval in Tris-EDTA, pH 9 and myeloperoxidase without antigen retrieval with anti-human myeloperoxidase antibody 1:5000 (A0398, Dako).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: Rheumatoid arthritis antigens homocitrulline and citrulline are generated by local myeloperoxidase and peptidyl arginine deiminases 2, 3 and 4 in rheumatoid nodule and synovial tissue

doi: 10.1186/s13075-016-1140-9

Figure Lengend Snippet: PAD3 ( c , g ) co-localization with citrulline- and homocitrulline-binding F95 antibody ( b ) and macrophage marker KP1 (CD68) ( g ) in the intact tissue adjacent to necrosis in rheumatoid nodule. PAD3 HIER was performed in pH 9 Tris-EDTA. PAD3 and F95 bound abundantly to the cytoplasm of cells adjacent to necrosis ( d ). PAD3 co-localized with KP1 in the cells adjacent to necrosis ( h ). Asterisk necrosis, double-headed arrow palisading histiocytes

Article Snippet: PAD2 enzyme was stained with anti-human PAD2 antibody 1:250 (ab16478, Abcam) and PAD3 with 1:50 (ab183209, Abcam) after heat-mediated antigen retrieval in Tris-EDTA, pH 9 and myeloperoxidase without antigen retrieval with anti-human myeloperoxidase antibody 1:5000 (A0398, Dako).

Techniques: Binding Assay, Marker