Journal: Oncotarget

Article Title: N -acetylcysteine negatively regulates Notch3 and its malignant signaling

doi: 10.18632/oncotarget.8806

Figure Lengend Snippet: A. Time- and dose-dependent inhibition of NAC on the intracellular domain of Notch3 (N3IC), but not Notch1 (N1IC). HeLa cells were treated with NAC (2-10 mM) for 0-24 h. B. Dose-dependent inhibition by NAC (0-10 mM, 6 h) on the protein expression of N3IC in HeLa cells. C. NAC treatment (5 mM, 0-12 h) reduces protein levels of N3IC and extracellular domain of Notch 3 (N3EC) but not full length Notch 3 precursor (N3FL) in HeLa cells. Densitometry quantifications of the protein bands were shown after normalization with their respective β-actin levels. Data are presented as means ± SE, n=3. *, p < 0.05 compared with their respective non-treated group.

Article Snippet: Mouse anti-Notch3 NECD antibody (H00004854-M01, Novus Biologicals, Littleton, CO, USA) recognizes the extracellular fragments and full length Notch3 precursor.

Techniques: Inhibition, Expressing

Journal: Oncotarget

Article Title: N -acetylcysteine negatively regulates Notch3 and its malignant signaling

doi: 10.18632/oncotarget.8806

Figure Lengend Snippet: A. NAC treatment (2-10 mM, 0-24 h) decreases Hes1 and HRT1 protein levels in HeLa cells. B. NAC treatment (0-10 mM for 6 h or 5 mM for 0-12 h) decreases Hes1 and HRT1 mRNA expression in HeLa cells. The mRNA expression of NAC-treated cells was normalized to that of non-treated cells whose value was set as 1. C. NAC treatment (0-10 mM, 12 h) inhibits Hes1 reporter activity in HeLa cells. The luciferase activity in NAC-treated cells was normalized to that of non-treated cells whose value was set as 1. D. Notch3 siRNA knockdown reduces Hes1 and HRT1 levels in HeLa cells. Protein levels were determined 2 days after siRNA transfection. siCtrl, scramble siRNA; siNotch3, Notch3 siRNA. Protein densitometry quantifications were shown after normalization with β-actin levels. Data are presented as means ± SE, n=3-4. *, p < 0.05 compared with their respective non-treated group.

Article Snippet: Mouse anti-Notch3 NECD antibody (H00004854-M01, Novus Biologicals, Littleton, CO, USA) recognizes the extracellular fragments and full length Notch3 precursor.

Techniques: Expressing, Activity Assay, Luciferase, Knockdown, Transfection

Journal: Oncotarget

Article Title: N -acetylcysteine negatively regulates Notch3 and its malignant signaling

doi: 10.18632/oncotarget.8806

Figure Lengend Snippet: A. Pre-treatment with a γ-secretase inhibitor, DAPT (20 μM, 30 min), had no effect on NAC-induced (5 mM, 0-12 h) decrease in N3IC protein expression in HeLa cells. B. NAC treatment (5 mM, 0-12 h) did not affect Notch3 mRNA expression in HeLa cells. C. Pre-treatment with NH 4 Cl (25 mM, 1 h), but not lactacystin (10 μM, 30 min), reversed NAC-induced (5 mM, 0-12 h) decrease of N3IC protein levels in HeLa cells. D. NAC treatment did not affect levels of exogenously expressed Notch3 active intracellular domain (N3ICD). HeLa cells were transfected with vectors expressing N3ICD or N3FL for 24 h, followed by treatment with NAC (5 mM, 0-12 h). E. Subcellular analysis of Notch3 protein levels following NAC treatment (5 mM, 6 h) in HeLa cells. Protein levels of N3FL, N3EC and N3IC in cytosolic, nuclear and membrane fractions were determined. Successful fractionation was evidenced by using the marker proteins GAPDH, cyclin B1, and Na + , K + -ATPase. N3FL, N3EC and N3IC denoted Notch3 full length, extracellular domain and intracellular domain, respectively. Protein densitometry quantifications were shown after normalization with β-actin (A, C, D) or their respective cellular compartment markers (E). Data are presented as means ± SE, n=3-4. *, p < 0.05 compared with their respective non-treated group.

Article Snippet: Mouse anti-Notch3 NECD antibody (H00004854-M01, Novus Biologicals, Littleton, CO, USA) recognizes the extracellular fragments and full length Notch3 precursor.

Techniques: Expressing, Transfection, Membrane, Fractionation, Marker

Journal: Oncotarget

Article Title: N -acetylcysteine negatively regulates Notch3 and its malignant signaling

doi: 10.18632/oncotarget.8806

Figure Lengend Snippet: N3ICD overexpression rescues NAC-induced inhibition of proliferation (A), migration (B), and invasion (C). A. Numbers of EV- and N3ICD-transfected cells were counted at 12-48 h after NAC treatment (0-10 mM, left panel). *, p < 0.05 compared with the EV-transfected cells within the same treatment and time point. B. Results of the wound healing assay (left panels) were expressed as the migration index (the distance migrated relative to the initial scraped gap) and that of EV-transfected cells without NAC treatment was set as 100% (middle panel). C. Cells per field on the insert membrane were imaged (left panels) and counted (middle panel). B and C: *, p < 0.05 compared with no NAC treatment; #, p < 0.05 compared with the EV-transfected cells within the same treatment. Percent rescue (A-C, right panels) after N3ICD expression was calculated by dividing the net change after NAC treatment in N3ICD-transfected cells by that in EV-transfected cells. Notch3 siRNA knockdown inhibits cell proliferation D. , migration E. , and invasion F. as assessed by the same approaches described above. Representative images for migration and invasion were shown. *, p < 0.05 compared with the siCtrl-transfected cells. All data are presented as mean ±SE, n=3. I, the initial seeded cell number. EV, empty vector; N3ICD, Notch3 active intracellular domain; siCtrl, scrambled siRNA; siNotch3, Notch3 siRNA.

Article Snippet: Mouse anti-Notch3 NECD antibody (H00004854-M01, Novus Biologicals, Littleton, CO, USA) recognizes the extracellular fragments and full length Notch3 precursor.

Techniques: Over Expression, Inhibition, Migration, Transfection, Wound Healing Assay, Membrane, Expressing, Knockdown, Plasmid Preparation

Journal: Oncotarget

Article Title: N -acetylcysteine negatively regulates Notch3 and its malignant signaling

doi: 10.18632/oncotarget.8806

Figure Lengend Snippet: A. NAC treatment (5 and 10 mM, 0-24 h) decreases N3IC protein levels in HCC1937 cells. Expression of exogenous N3ICD rescues NAC-induced inhibition of proliferation B. , migration C. , and invasion D. , and Notch3 siRNA knockdown inhibits proliferation E. , migration F. , and invasion G. in HCC1937 cells. Quantifications, sample size, statistics, and abbreviations for protein levels, proliferation, migration, and invasion assays were as described in Figure & legends.

Article Snippet: Mouse anti-Notch3 NECD antibody (H00004854-M01, Novus Biologicals, Littleton, CO, USA) recognizes the extracellular fragments and full length Notch3 precursor.

Techniques: Expressing, Inhibition, Migration, Knockdown

Journal: PLoS ONE

Article Title: Necl-4/SynCAM-4 Is Expressed in Myelinating Oligodendrocytes but Not Required for Axonal Myelination

doi: 10.1371/journal.pone.0064264

Figure Lengend Snippet: A. Western immuneblotting of brain tissues (from P10 wild-type and Necl-4 homozygous mice) with anti-Necl-1, anti-Necl-2 and anti-Necl-3 antibodies. β-actin was the internal control. B. Statistical analysis on the relative expression level of Necl1, Necl2 and Necl3 with Student's t-test (n = 3. Necl1, p = 0.70. Necl2, p = 0.72. Necl3, p = 0.69). Error bar, standard deviation.

Article Snippet: 30 mg protein from control and mutant tissues was loaded for SDS-PAGE electrophoresis and subsequently detected with anti-Necl-1 (developed in Peking Union Medical University), anti-Necl-2 (Proteintech, Cat# 14335-1-AP), anti-Necl-3 (Abcam Inc, Cat# ab133393) and anti-Necl-4 (UC Davis/NIH NeuroMab Facility, Cat#73-247), and mouse anti-β-actin (Sigma, Cat# A5316) antibodies according to the standard protocol.

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Frontiers in Cellular Neuroscience

Article Title: Secretion of Mast Cell Inflammatory Mediators Is Enhanced by CADM1-Dependent Adhesion to Sensory Neurons

doi: 10.3389/fncel.2019.00262

Figure Lengend Snippet: CADM1 is expressed in BMMCs and sensory neurons. (A) Immunoblot of lysates prepared from DRG and BMMC mono-cultures probed with anti-CADM1 and anti-α-Tubulin, as a loading control. The m.w. scale is shown to the right of the blot. (B) Flow cytometric analysis of surface (left) and total CADM1 expression in Triton X-100-permeablized BMMCs (right). BMMCs were double labeled with conjugated c-Kit and CADM1 Abs. BMMCs were gated as c-kit+ CADM1high cell subset and c-kit+ CADM1medium cell subset. Numbers in plots are the percentages of cells in the indicated gate. (C) Confocal immunofluorescence images of c-Kit (red), CADM1 (green), and DAPI (blue) and merged image of mono-cultured BMMCs. Scale bar represents 10 μm. (D,E) Confocal immunofluorescence images of β-Tubulin (red), CADM1 (green) and DAPI (blue) in mono-cultured DRGs. White arrows indicate CADM1 immunoreactive neurites. Note that non-neuronal cells, observed as DAPI positive, β-Tubulin negative are also negative for CADM1 (arrowheads). Scale bar represents 20 μm.

Article Snippet: Then, the membrane was probed using rabbit anti-mouse CADM1 antibody (1:300, Santa Cruz) and mouse anti-GAPDH (1:5000, Thermo Fisher Scientific) and visualized using Li-cor system, Goat IRDye 800 anti-Rabbit 1:5000 and Goat IRDye 700 anti-Mouse 1:5000.

Techniques: Western Blot, Control, Expressing, Labeling, Immunofluorescence, Cell Culture

Journal: Frontiers in Cellular Neuroscience

Article Title: Secretion of Mast Cell Inflammatory Mediators Is Enhanced by CADM1-Dependent Adhesion to Sensory Neurons

doi: 10.3389/fncel.2019.00262

Figure Lengend Snippet: CADM1 is enriched at mast cell-sensory neurite contact sites. Representative immunofluorescent images of BMMCs co-cultured with DRG neurons for 24 h. In panel (A) individual and overlaid images are shown from a neurite with two adherent cells. β-Tubulin (red) is used as the neuronal marker, DAPI (blue) nuclear marker labels all cells in the culture, while FcεRI (yellow) specifically labels BMMCs. CADM1 (green) stained positive in the sensory neuron neurite and BMMC, but is absent from the attached non-neuronal, non-mast cell. (B,C) Representative individual and overlaid immunofluorescent images of CADM1 (green), β-Tubulin (red) and DAPI (blue) staining of different co-cultures. Increased CADM1 fluorescent intensity at contact sites (white arrows) between BMMCs and neurites (red).

Article Snippet: Then, the membrane was probed using rabbit anti-mouse CADM1 antibody (1:300, Santa Cruz) and mouse anti-GAPDH (1:5000, Thermo Fisher Scientific) and visualized using Li-cor system, Goat IRDye 800 anti-Rabbit 1:5000 and Goat IRDye 700 anti-Mouse 1:5000.

Techniques: Cell Culture, Marker, Staining

Journal: Frontiers in Cellular Neuroscience

Article Title: Secretion of Mast Cell Inflammatory Mediators Is Enhanced by CADM1-Dependent Adhesion to Sensory Neurons

doi: 10.3389/fncel.2019.00262

Figure Lengend Snippet: Adhesion of mast cells to sensory neurons is attenuated by a CADM1 blocking peptide. (A) Superimposed bright-field and fluorescent image of live calcein-labeled, adherent BMMCs (green) co-cultured with DRG neurons (unlabeled cells) for 2 h. Non-adherent cells have been removed by washing and centrifugation of the plate. (B) Image of calcein-labeled BMMCs plated in parallel into matrigel-coated wells devoid of neurons and subjected to the same washing and centrifugation procedure. (C) BMMC adhesion quantified from calcein-fluorescence remaining in wells after washing and centrifugation expressed as a percentage of total well fluorescence measured prior to washing procedure. Data shown as mean ± SEM from N = 3. Each done in duplicate. Data were analyzed using a two-tailed paired t -test ∗∗ p < 0.01. (D) Concentration-dependent inhibition of mast cell adhesion to DRG measured with a CADM1 blocking peptide. Percentage of adherent BMMC was calculated using the calcein adhesion assay. Each condition was done in duplicate, on N = 3 cultures. Each point represents the mean ± SEM. One-way ANOVA followed by Turkey’s multiple comparison post-test was performed. ∗∗ denotes p < 0.01 and ∗∗∗ p < 0.001 compared to the percentage of BMMC adhesion in the absence of CADM1 blocking peptide.

Article Snippet: Then, the membrane was probed using rabbit anti-mouse CADM1 antibody (1:300, Santa Cruz) and mouse anti-GAPDH (1:5000, Thermo Fisher Scientific) and visualized using Li-cor system, Goat IRDye 800 anti-Rabbit 1:5000 and Goat IRDye 700 anti-Mouse 1:5000.

Techniques: Blocking Assay, Labeling, Cell Culture, Centrifugation, Fluorescence, Two Tailed Test, Concentration Assay, Inhibition, Cell Adhesion Assay, Comparison

Journal: Frontiers in Cellular Neuroscience

Article Title: Secretion of Mast Cell Inflammatory Mediators Is Enhanced by CADM1-Dependent Adhesion to Sensory Neurons

doi: 10.3389/fncel.2019.00262

Figure Lengend Snippet: Knockdown of CADM1 in mast cells inhibits their adhesion to sensory neurons. (A) Western blot analysis of CADM1 expression in BMMCs transfected with the indicated ShRNA 48 h previously. Expression from un-transfected control cells examined in parallel are also shown (lane labeled BMMC). GAPDH was used as loading control. Data are representative of three independent experiments. (B) Adherence of BMMCs transfected with CADM1 ShRNA or scrambled control to neurons tested using the calcein assay. BMMC adhesion in the absence of DRG neurons is shown as a control. CADM1 knockdown significantly reduced BMMC adhesion to sensory neurons ( N = 3). Each bar represents the mean ± SEM. One-way ANOVA followed by Turkey’s multiple comparison post-test was performed. ∗∗∗ denotes p < 0.001.

Article Snippet: Then, the membrane was probed using rabbit anti-mouse CADM1 antibody (1:300, Santa Cruz) and mouse anti-GAPDH (1:5000, Thermo Fisher Scientific) and visualized using Li-cor system, Goat IRDye 800 anti-Rabbit 1:5000 and Goat IRDye 700 anti-Mouse 1:5000.

Techniques: Knockdown, Western Blot, Expressing, Transfection, shRNA, Control, Labeling, Comparison

Journal: Frontiers in Cellular Neuroscience

Article Title: Secretion of Mast Cell Inflammatory Mediators Is Enhanced by CADM1-Dependent Adhesion to Sensory Neurons

doi: 10.3389/fncel.2019.00262

Figure Lengend Snippet: CADM1 is necessary for mast-cell sensor neuron cross-talk. BMMCs pre-sensitized with anti-DNP IGE were co-cultured with DRG for 24 h in the presence or absence of 10 μg/ml CADM1 blocking peptide before degranulation was quantified using the β-Hex assay. Data were normalized to degranulation measurements made from BMMC mono-cultures set up in parallel, and the fold-change in basal Ag-Indep degranulation (A) or DNP (10 ng/ml), Ag-activated degranulation (B) calculated and compared by two-tailed paired t -test, ∗ denotes p < 0.05, ∗∗ p < 0.01. Each bar represents the mean ± SEM ( N = 3). (C) Untransfected (WT BMMC), CADM1-ShRNA or scramble-transfected BMMCs pre-sensitized with anti-DNP IGE were co-cultured with DRG for 24 h before degranulation was quantified using the β-Hex assay. Fold change of Ag-Independent degranulation (C) and DNP (10 ng/ml) Ag-activated degranulation (D) were calculated and compared by one-way ANOVA followed by Turkey’s multiple comparison post-test, ∗∗∗ denotes p < 0.001 compared to wt BMMC-DRG co-culture. Each bar represents the mean ± SEM ( N = 3).

Article Snippet: Then, the membrane was probed using rabbit anti-mouse CADM1 antibody (1:300, Santa Cruz) and mouse anti-GAPDH (1:5000, Thermo Fisher Scientific) and visualized using Li-cor system, Goat IRDye 800 anti-Rabbit 1:5000 and Goat IRDye 700 anti-Mouse 1:5000.

Techniques: Cell Culture, Blocking Assay, Two Tailed Test, shRNA, Transfection, Comparison, Co-Culture Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: Secretion of Mast Cell Inflammatory Mediators Is Enhanced by CADM1-Dependent Adhesion to Sensory Neurons

doi: 10.3389/fncel.2019.00262

Figure Lengend Snippet: Sensory neuron potentiation of antigen-stimulated cytokine secretion by mast cells is dependent on CADM1. BMMCs pre-sensitized with anti-DNP IGE were cultured alone or with DRG for 24 h. (A) Secreted IL-6 and (B) TNFα were quantified by ELISA for cells stimulated in parallel with the antigen DNP (10 ng/ml) for 6 h or vehicle control. Data shown are mean ± SEM of N = 3, each performed in duplicate. ∗ p < 0.05 compared to stimulated BMMC alone. Data were analyzed using two-tailed paired t -test. (C) BMMC mono-culture and untransfected WT, CADM1-knockdown or scramble-transfected BMMCs were co-cultured with DRG for 24 h, prior to being activated with DNP (10 ng/ml) for 6 h. IL-6 was measured from supernatants and compared by one-way ANOVA followed by Turkey’s multiple comparison post-test, ∗ denotes p < 0.05 compared to wt BMMC-DRG co-culture. Each bar represents the mean ± SEM ( N = 3).

Article Snippet: Then, the membrane was probed using rabbit anti-mouse CADM1 antibody (1:300, Santa Cruz) and mouse anti-GAPDH (1:5000, Thermo Fisher Scientific) and visualized using Li-cor system, Goat IRDye 800 anti-Rabbit 1:5000 and Goat IRDye 700 anti-Mouse 1:5000.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Control, Two Tailed Test, Knockdown, Transfection, Comparison, Co-Culture Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Antiviral Cytotoxic T Lymphocytes in Vaginal Mucosa of Simian Immunodeficiency Virus-Infected-Rhesus Macaques 1

doi:

Figure Lengend Snippet: A single, representative frozen section of vaginal mucosa from a chronically SIV-infected monkey was stained with three mAbs. A, CD2+ T cells (green) in vaginal epithelium. B, CD8+ cells (red) in same tissue section. C, Immunofluorescence double staining of CD2+/CD8+ cells in same section of vaginal epithelium. Note that all CD2+ T cells in vaginal epithelium are CD8+. Double-labeled cells are yellow, some cells (arrows) have variably distinct areas of red (CD8 expression) and yellow (colocalization of CD2 and CD8) staining or green (CD2 expression) and yellow (colocalization of CD2 and CD8) staining. D, TIA-1+ cytoplasmic granules (blue) in CD8+ T cells in same section of vaginal epithelium. E, Immunofluorescence triple staining of CD2+/CD8+/TIA-1+ T cells in vaginal epithelium. This figure demonstrates that all CD2+, CD8+ T cells in vaginal epithelium are TIA-1+. F, A computer-labeled image to clarify anatomic components in A-E. Original magnification: ×630.

Article Snippet: CD8 + cells were positively selected from the cell suspension using two rounds of immunomagnetic bead isolation (M450 anti-CD8 Dynabeads; Dynal, Great Neck, NY).

Techniques: Infection, Staining, Immunofluorescence, Double Staining, Labeling, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Antiviral Cytotoxic T Lymphocytes in Vaginal Mucosa of Simian Immunodeficiency Virus-Infected-Rhesus Macaques 1

doi:

Figure Lengend Snippet: A frozen section of vagina (serial section of slide in Fig. 2). A-C, Photographs of a single representative cryostat section stained with two mAbs. A, Cells expressing TCR β-chain (green) in vaginal mucosa. B, CD8+ cells (red) in same section of vaginal mucosa. C, Coexpression of CD8 and TCR β-chain in vaginal epithelial T cells. Double-labeled cells are yellow. This figure demonstrates expression of TCR β-chain by all CD8+ T cells in vaginal epithelium. Original magnification: ×630.

Article Snippet: CD8 + cells were positively selected from the cell suspension using two rounds of immunomagnetic bead isolation (M450 anti-CD8 Dynabeads; Dynal, Great Neck, NY).

Techniques: Staining, Expressing, Labeling

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Antiviral Cytotoxic T Lymphocytes in Vaginal Mucosa of Simian Immunodeficiency Virus-Infected-Rhesus Macaques 1

doi:

Figure Lengend Snippet: Comparison of SIV-specific pCTL frequencies in vaginal epithelium from a chronically SIV-infected rhesus macaque (A), and an acutely SIV-infected rhesus macaque (B). Vaginal epithelial CD8+ lymphocytes were stimulated in culture for 14 days and then analyzed for cytotoxic activity against autologous lymphoblastoid cells infected with vvWR, vvgag (squares), and vvenv (triangles). Percentage of negative wells is plotted against number of CD8+ T cells per well, and frequency is extrapolated from × intercept of regression line with 37%.

Article Snippet: CD8 + cells were positively selected from the cell suspension using two rounds of immunomagnetic bead isolation (M450 anti-CD8 Dynabeads; Dynal, Great Neck, NY).

Techniques: Infection, Activity Assay

Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

Article Title: Overlapping morphological, immunohistochemical and genetic features of superficial CD34-positive fibroblastic tumor and PRDM10-rearranged soft tissue tumor.

doi: 10.1038/s41379-021-00991-8

Figure Lengend Snippet: Fig. 2 Immunoreactivity for CADM3. A, B Moderate to strong immunoreactivity for CADM3 was seen in 41 of 43 SCD34FT/PRDM10- rearranged soft tissue tumors depicting classical morphology (case 9). C, D Immunoreactivity was also noted in cases with pronounced edema (case 42); E, F and in cases with predominant spindle cell morphology (case 18).

Article Snippet: For IHC, antigen retrieval was performed on 3 μm sections from formalinfixed paraffin-embedded tissue in pH 9 Tris/EDTA buffer retrieval solution (Agilent, Santa Clara, CA) at 68 °C for 16 h. For the detection of CADM3, the primary mouse monoclonal CADM3 antibody raised against human CADM3 Pro21-Tyr329 (R&D Systems, Minneapolis, MN, USA; clone #730004,) was used (1:100 dilution).

Techniques:

Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

Article Title: Overlapping morphological, immunohistochemical and genetic features of superficial CD34-positive fibroblastic tumor and PRDM10-rearranged soft tissue tumor.

doi: 10.1038/s41379-021-00991-8

Figure Lengend Snippet: Fig. 3 Patterns of immunoreactivity for CADM3 in SCD34FT/PRDM10-rearranged soft tissue tumors. A 41 of 43 tumors showed strong (left) or moderate (right) immunoreactivity, with some geographical variety (cases 29, 33). B Except for peripheral nerves (arrowheads), there was no staining of non-tumoral soft tissue components (case 39, re-excision specimen). C Immunoreactivity was granular, with membranous accentuation (case 15). D Cytoplasmic positivity was slightly weaker and granular (case 39). E, F Granular cell change in tumor cells showed more pronounced immunoreactivity contrasting with surrounding spindle cell areas (case 23).

Article Snippet: For IHC, antigen retrieval was performed on 3 μm sections from formalinfixed paraffin-embedded tissue in pH 9 Tris/EDTA buffer retrieval solution (Agilent, Santa Clara, CA) at 68 °C for 16 h. For the detection of CADM3, the primary mouse monoclonal CADM3 antibody raised against human CADM3 Pro21-Tyr329 (R&D Systems, Minneapolis, MN, USA; clone #730004,) was used (1:100 dilution).

Techniques: Staining

Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

Article Title: Overlapping morphological, immunohistochemical and genetic features of superficial CD34-positive fibroblastic tumor and PRDM10-rearranged soft tissue tumor.

doi: 10.1038/s41379-021-00991-8

Figure Lengend Snippet: Fig. 4 AE1/AE3 immunoreactivity in SCD34FT/PRDM10-rearranged soft tissue tumors and CADM3 in other soft tissue tumors. A, B AE1/ AE3 immunoreactivity in SCD34FT/PRDM10-rearranged soft tissue tumors varied from diffuse (case 40) to very focal (case 15) to negative. C, D Focal immunoreactivity for CADM3 was noted in 3 out of 26 myxofibromsarcomas tested. E, F Four out of 10 pleomorphic liposarcomas showed focal positivity for CADM3.

Article Snippet: For IHC, antigen retrieval was performed on 3 μm sections from formalinfixed paraffin-embedded tissue in pH 9 Tris/EDTA buffer retrieval solution (Agilent, Santa Clara, CA) at 68 °C for 16 h. For the detection of CADM3, the primary mouse monoclonal CADM3 antibody raised against human CADM3 Pro21-Tyr329 (R&D Systems, Minneapolis, MN, USA; clone #730004,) was used (1:100 dilution).

Techniques:

Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

Article Title: Overlapping morphological, immunohistochemical and genetic features of superficial CD34-positive fibroblastic tumor and PRDM10-rearranged soft tissue tumor.

doi: 10.1038/s41379-021-00991-8

Figure Lengend Snippet: Fig. 5 CADM3 immunoreactivity in tumors containing Schwann cells and nerves. A There was membranous immunoreactivity in schwannomas, B and nerve sheath myxomas. C, D Granular cell tumors showed granular cytoplasmic positivity. E In neurofibromas, CADM3 stained intralesional myelinated and unmyelinated axons but no lesional cells. F In longitudinal sections of myelinated peripheral nerves, crosstie-like immunreactivity for CADM3 highlighted Schmidt-Lanterman incisures (arrowheads).

Article Snippet: For IHC, antigen retrieval was performed on 3 μm sections from formalinfixed paraffin-embedded tissue in pH 9 Tris/EDTA buffer retrieval solution (Agilent, Santa Clara, CA) at 68 °C for 16 h. For the detection of CADM3, the primary mouse monoclonal CADM3 antibody raised against human CADM3 Pro21-Tyr329 (R&D Systems, Minneapolis, MN, USA; clone #730004,) was used (1:100 dilution).

Techniques: Staining

Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

Article Title: Overlapping morphological, immunohistochemical and genetic features of superficial CD34-positive fibroblastic tumor and PRDM10-rearranged soft tissue tumor.

doi: 10.1038/s41379-021-00991-8

Figure Lengend Snippet: Fig. 6 Overview of cases with diagnoses, immunhistochemical and molecular findings. Of 18 cases with PRDM10 rearrangements, 7 were originally classified as undifferentiated pleomorphic sarcoma (5) or liposarcoma (2) and 11 were classified as SCD34FT. All tumors with PRDM10 rearrangement showed immunoreactivity for CADM3.

Article Snippet: For IHC, antigen retrieval was performed on 3 μm sections from formalinfixed paraffin-embedded tissue in pH 9 Tris/EDTA buffer retrieval solution (Agilent, Santa Clara, CA) at 68 °C for 16 h. For the detection of CADM3, the primary mouse monoclonal CADM3 antibody raised against human CADM3 Pro21-Tyr329 (R&D Systems, Minneapolis, MN, USA; clone #730004,) was used (1:100 dilution).

Techniques: