Journal: Pathology and Oncology Research

Article Title: High prevalence of Wilms tumor 1 expression in multiple myeloma and plasmacytoma: A cohort of 142 Asian patients’ samples

doi: 10.3389/pore.2023.1610844

Figure Lengend Snippet: WT1 protein expression in tissues at different stages of multiple myeloma (MM): Rates of WT1 protein expression in MM tissues at different clinical settings (A) , Rates of cytoplasmic and nuclear WT1 staining in different tissue samples (B) , Histology score (H-score) of WT1 cytoplasmic staining (C) , and nuclear staining (D) in different myeloma samples at first diagnosis or relapse stage (* p < 0.05, ** p < 0.01).

Article Snippet: Prediluted 1:500 mouse anti-WT1 antibody (clone 6F-H2; Cell Marque) was incubated for 1 h at 36°C.

Techniques: Expressing, Staining, Biomarker Discovery

Journal: Pathology and Oncology Research

Article Title: High prevalence of Wilms tumor 1 expression in multiple myeloma and plasmacytoma: A cohort of 142 Asian patients’ samples

doi: 10.3389/pore.2023.1610844

Figure Lengend Snippet: Immunohistochemistry (IHC) staining of WT1 protein: (A–D) show a representative cytoplasmic staining with different intensities in samples: Negative staining (A) , + (B) , ++ (C) and +++ (D) ; (E–H) show a representative nuclear staining with different intensities in samples: Negative staining (E) , + (F) , ++ (G) and +++ (H) .

Article Snippet: Prediluted 1:500 mouse anti-WT1 antibody (clone 6F-H2; Cell Marque) was incubated for 1 h at 36°C.

Techniques: Immunohistochemistry, Staining, Negative Staining

Journal: Pathology and Oncology Research

Article Title: High prevalence of Wilms tumor 1 expression in multiple myeloma and plasmacytoma: A cohort of 142 Asian patients’ samples

doi: 10.3389/pore.2023.1610844

Figure Lengend Snippet: Characteristics of WT1 positivity in samples.

Article Snippet: Prediluted 1:500 mouse anti-WT1 antibody (clone 6F-H2; Cell Marque) was incubated for 1 h at 36°C.

Techniques: Staining, Isolation

Journal: Scientific Reports

Article Title: Reporter cell assay for human CD33 validated by specific antibodies and human iPSC-derived microglia

doi: 10.1038/s41598-021-92434-2

Figure Lengend Snippet: CD33-specific and control antibodies.

Article Snippet: , P67.6 F(ab) , Human , Mouse , Sanofi , F(ab) fragment of P67.6 , All.

Techniques: Imaging

Journal: Scientific Reports

Article Title: Reporter cell assay for human CD33 validated by specific antibodies and human iPSC-derived microglia

doi: 10.1038/s41598-021-92434-2

Figure Lengend Snippet: Flow cytometric analysis of CD33 surface expression. ( a ) Schematic drawing of the CD33-DAP12 constructs. Both, the full CD33 ecto-domain (CD33M) and the ecto-domain lacking the sialic acid binding domain (CD33 ∆E2 ) were fused to TYROBP/DAP12. CD33 ∆E2 can be identified by binding of the antibody clone 1c7/1 (blue) but not WM53 or P67.6 (red), whereas all three antibody clones can bind CD33M. ( b ) The CD33-DAP12 and CD33-DAP12-GCaMP6m cells were stained for CD33 surface expression with the antibody clones 1c7/1, WM53 and P67.6. A representative flow cytometry histogram plot for the CD33M-DAP12-GCaMP6m cells is shown (left side). All three tested antibodies were able to stain full-length CD33 on the cell surface. Expression of variant 2 CD33 from CD33 ∆E2 -DAP12 and CD33 ∆E2 -DAP12-GCaMP6m cells was only detected by antibody clone 1c7/1. A representative flow cytometry histogram plot for the CD33 ∆E2 -DAP12-GCaMP6m cells is shown (right side). ( c ) Quantification of CD33 staining showed a high percentage of CD33 expressing cells in the CD33M-DAP12-GCaMP6m line for all three tested antibody clones but only the CD33 antibody clone 1c7/1 was able to detect CD33 in CD33 ∆E2 -DAP12-GCaMP6m expressing cells. The antibody clones WM53 and P67.6 did not show any staining of CD33 ∆E2 -DAP12 expressing cells. ( d ) Quantification of CD33 staining revealed a high percentage of cells in the CD33M-DAP12 line expressed CD33, and was detected by all three antibody clones. CD33 in CD33 ΔE2 -DAP12 expressing cells was only detected by antibody clone 1c7/1. Data are shown as mean + SEM of three to five independent experiments; *** p ≤ 0.001 compared to Secondary Control determined by Welch ANOVA followed by Games-Howell post hoc test.

Article Snippet: , P67.6 F(ab) , Human , Mouse , Sanofi , F(ab) fragment of P67.6 , All.

Techniques: Expressing, Construct, Binding Assay, Clone Assay, Staining, Flow Cytometry, Variant Assay

Journal: Scientific Reports

Article Title: Reporter cell assay for human CD33 validated by specific antibodies and human iPSC-derived microglia

doi: 10.1038/s41598-021-92434-2

Figure Lengend Snippet: pSYK detection in CD33M-DAP12 cell lines. ( a ) pSYK detection in CD33M-DAP12 reporter cells treated with CD33-specific antibodies. Addition of CD33 antibodies P67.6 and 1c7/1 resulted in increased pSYK levels, whilst P67.6 F(ab), WM53 as well as the different isotype IgG1 control antibodies did not show any change in endogenous pSYK levels. ( b ) 1c7/1 and P67.6 dose–response curve in CD33M-DAP12 reporter cells. Addition of CD33 antibody clones 1c7/1 and P67.6 resulted in an increase in endogenous pSYK levels measured 30 min after the treatment. Data are presented as mean ± or + SD; ** p ≤ 0.01 compared to untreated determined by one-way ANOVA analysis followed by Dunnett’s post hoc test.

Article Snippet: , P67.6 F(ab) , Human , Mouse , Sanofi , F(ab) fragment of P67.6 , All.

Techniques: Clone Assay

Journal: Scientific Reports

Article Title: Reporter cell assay for human CD33 validated by specific antibodies and human iPSC-derived microglia

doi: 10.1038/s41598-021-92434-2

Figure Lengend Snippet: Calcium imaging in CD33-DAP12-GCaMP6m reporter cell lines. ( a ) Schematic time line of image acquisition and compound handling. ( b , c ) Calcium imaging analyzed as ΔF/F(t) in CD33-DAP12-GCaMP6m lines. Addition 100 µM dATP led to a strong increase in intracellular calcium levels in both cell lines, CD33M- and CD33 ∆E2 -DAP12-GCaMP6m, with a peak at around 20–25 s. The CD33 antibody clones 1c7/1 and P67.6 evoked a selective intracellular calcium response only in CD33M-DAP12-GCaMP6m cells. The CD33 antibody clones WM53 and P67.6 F(ab) as well as the isotype IgG1/F(ab’)2 antibodies did not show a change in intracellular calcium levels. ( d , e ) The area under the curve as well as the maximum ΔF/F(t) signal calculated from independent experiments showed a significant increase in dATP treated samples in both CD33-DAP12-GCaMP6m lines and a selective increase in CD33M-DAP12-GCaMP6m expressing cells if treated with the CD33 antibody clone P67.6 or 1c7/1. Data are presented mean + SEM; n = 3–6; *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05 compared to 10 µg/ml IgG1 determined by Welch ANOVA followed by Games-Howell post hoc test.

Article Snippet: , P67.6 F(ab) , Human , Mouse , Sanofi , F(ab) fragment of P67.6 , All.

Techniques: Imaging, Clone Assay, Expressing

Journal: Scientific Reports

Article Title: Reporter cell assay for human CD33 validated by specific antibodies and human iPSC-derived microglia

doi: 10.1038/s41598-021-92434-2

Figure Lengend Snippet: Activation of endogenous CD33 in iPSdMiG by CD33 agonistic antibodies. ( a ) pSYK analysis in TREM2 + DAP12 reporter cells. Addition of anti-TREM2 antibody AF1828 resulted in an increase in endogenous pSYK levels measured 30 min after the treatment only in TREM2 + DAP12 but not DAP12 expressing control reporter cells. Data are presented mean ± SD. ( b ) CD33 antibodies P67.6 and 1c7/1 were able to decrease the increased pSYK/tSYK levels triggered by TREM2 activation in WT iPSdMiG (left). In CD33 −/− (middle) and CD33 ΔE2 (right) iPSdMiG none of the tested antibodies was able to modulate pSYK/tSYK levels after TREM2 activation. Data are presented mean + SEM; n = 3–6; ** p ≤ 0.01 compared to IgG1 (anti-CD33 Ctrl) determined by Welch ANOVA followed by Games-Howell post hoc test. ( c ) CD33 antibodies P67.6 and 1c7/1 decreased the phagocytic uptake of pHrodo S. aureus BioParticles in WT iPSdMiG (left). In CD33 −/− (middle) and CD33 ΔE2 (right) iPSdMiG none of the tested antibodies was able to modulate pHrodo S. aureus BioParticle phagocytosis. Data are presented mean + SEM; n = 3; ** p ≤ 0.01 and * p ≤ 0.05 compared to IgG1 determined by ANOVA followed by Dunnett’s post hoc test.

Article Snippet: , P67.6 F(ab) , Human , Mouse , Sanofi , F(ab) fragment of P67.6 , All.

Techniques: Activation Assay, Expressing

Journal: Oncology Letters

Article Title: Spindle cell sarcoma with KIAA1549-BRAF resembling infantile fibrosarcoma morphologically: A case report and literature review

doi: 10.3892/ol.2022.13572

Figure Lengend Snippet: IHC staining (magnification, ×400). (A) Weak positive expression of desmin. (B) Cytoplasmic expression of WT-1. Negative IHC staining results for (C) pan-TRK, (D) S-100, (E) SMA, (F) CD34, (G) CD99, (H) NKX2.2, (I) MyoD1 and (J) pan-cytokeratin. IHC, immunohistochemistry; WT-1, wilms tumor gene 1; pan-TRK, pan-tropomyosin receptor kinase; SMA, smooth muscle actin; NKX2.2, NK2 homeobox 2; MyoD1, myogenic differentiation 1.

Article Snippet: Immunohistochemical staining was performed on 4-µm-thick sections using a fully automated systems [Bench Mark GX System (Roche, Rotkreuz, Switzerland) or Leica Bond-max (Leica Biosystems, Buffalo Grove, IL, USA)] and the following primary antibodies; desmin (clone D33; prediluted) (IR606; Dako, Calpinteria, CA, USA), Wilms tumor gene 1 (WT-1) (6F-H2; dilution 1:1) (41386; NICHIREI, Osaka, Japan), pan-TRK [VENTANA Pan-TRK (EPR17341) Assay] (790–7026; Roche), S-100 (polyclonal; dilution 1:2,000) (Z3011; Dako), SMA (clone 1A4; dilution 1:300) (IR611; Dako), CD34 (clone NU-4A1; dilution 1:4) (413111; NICHIREI, Osaka Japan), CD99 (clone O13; prediluted) (790–4452; Roche), NK2 homeobox 2(NKX2.2) (rabbit polyclonal; dilution 1:50) (NBP1-82554; Novus Biologicals, Littleton, CO, USA), myogenic differentiation 1 (MyoD1) (mouse monoclonal; dilution 1:250) (ab133627, abcam, USA), and pan-cytokeratin (clone AE1/AE3; dilution 1:100) (IR053; Dako).

Techniques: Immunohistochemistry, Expressing, Wilms Tumor Assay