Journal: Cancer Research Communications

Article Title: SM08502-Mediated β-Catenin Repression Synergizes with Olaparib to Inhibit Tumor Progression

doi: 10.1158/2767-9764.CRC-25-0267

Figure Lengend Snippet: SM08502 and olaparib remodel the immune TME. A, ID8-OR tumors from mice treated with control, olaparib, SM08502, or in combination (ola + SM) were used for multispectral IHC. Five tumor regions were selected from each mouse ( n = 5). B, Assessment of apoptosis via cleaved caspase 3 (CC3)-positive tumor (WT1+) cells. C, Assessment of proliferation Ki67-positive cells. D, Assessment of DNA damage via γH2Ax. E, CD3 + (T cells) and ( F ) PD-1+CD3 + (PD-1–expressing T cells). G, Tumor-associated macrophages (F4/80+). H, PD-L1+ expression in F4/80+ cells. Outlier analysis via ROUT (Q = 1%) was conducted on all graphs. Error bars, SEM. Statistical test, one-way ANOVA with Tukey multiple comparison correction.

Article Snippet: Murine tumor sections were sequentially stained with antibodies specific for γH2Ax (1:400, Opal 620, Abcam, cat. #ab2893, RRID: AB_303388), Pd-l1 (1:100, Opal 690, R and D Systems, cat. #AF1019, RRID: AB_354540), cleaved caspase 3 (1:100, Opal 540, Cell Signaling Technology, cat. #9661, RRID: AB_2341188), Cd3 (1:50, Opal 520, Cell Signaling Technology, cat. #99940, RRID: AB_2755035), Wt1 (1:400, Opal 650, Novus, cat. #NB110-60011B, RRID: AB_1849479), Pd-1 (1:150, Opal 570, Cell Signaling Technology, cat. #84651, RRID: AB_2800041), Ki67 (1:400, Opal 480, Thermo Fisher Scientific, cat. #RM-9106, RRID: AB_2341197), and F4/80 (1:50, Opal 780, Cell Signaling Technology, cat. #30325, RRID: AB_2798990).

Techniques: Control, Expressing, Comparison

Journal: Stem Cell Research & Therapy

Article Title: Amniotic fluid stem cells ameliorate cisplatin-induced acute renal failure through induction of autophagy and inhibition of apoptosis

doi: 10.1186/s13287-019-1476-6

Figure Lengend Snippet: List of antibodies with their dilutions, sources, and catalog numbers

Article Snippet: Mouse monoclonal WT1 (IgG clone #6F-H2) , 1:100 (FCM) , My BioSource, CA, USA.

Techniques: Plasmid Preparation

Journal: Stem Cell Research & Therapy

Article Title: Amniotic fluid stem cells ameliorate cisplatin-induced acute renal failure through induction of autophagy and inhibition of apoptosis

doi: 10.1186/s13287-019-1476-6

Figure Lengend Snippet: Morphology and phenotypic characterization of AFSC. a Representative photomicrographs of amniotic fluid stem cells (AFSC) in culture showing spindle-shaped morphology in passage 3 (scale bar: 100 μm). b Phenotypic characterization of AFSC by flow cytometry showing the expression of cell-surface markers, viz, CD73, CD90, CD105, MHC Class II, and CD45, and intracellular renal progenitor markers, viz, WT1, SIX2, and PAX2 (green or red lines detected with FITC- and PE-conjugated antibodies, respectively, and black lines represent isotype controls). PE, phycoerythrin; FITC, fluorescein isothiocyanate

Article Snippet: Mouse monoclonal WT1 (IgG clone #6F-H2) , 1:100 (FCM) , My BioSource, CA, USA.

Techniques: Flow Cytometry, Expressing

Journal: Leukemia

Article Title: GATA-1 and GATA-2 binding to 3' enhancer of WT1 gene is essential for its transcription in acute leukemia and solid tumor cell lines.

doi: 10.1038/leu.2009.13

Figure Lengend Snippet: Figure 1 WT1, GATA-1 and GATA-2 mRNA in leukemia cell lines (a) and solid tumor cell lines (b). Relative WT1, GATA-1 and GATA-2 mRNA levels were examined by a quantitative reverse transcriptase-PCR. The internal control was GAPDH. The asterisks denote below the detection limit. The lower panels (c) show the correlation between WT1 and GATA-1, WT1 and GATA-2, and GATA-1 and GATA-2, respectively.

Article Snippet: Western blotting of WT1, GATA-1, GATA-2 and Sp1 was carried out using the anti-WT1 antibody (clone 6F-H2, Dakocytomation, CA, USA), anti-GATA-1 (sc-265), anti-GATA-2 (sc-9008) and anti-Sp1 (sc-59) antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), respectively.

Techniques: Reverse Transcription, Control

Journal: Leukemia

Article Title: GATA-1 and GATA-2 binding to 3' enhancer of WT1 gene is essential for its transcription in acute leukemia and solid tumor cell lines.

doi: 10.1038/leu.2009.13

Figure Lengend Snippet: Figure 2 WT1, GATA-1, GATA-2 and Sp1 protein levels of representative cell lines. Left: Representative leukemia cell lines with high (K562, HEL, KU812F and HL60 cells) and low WT1 mRNAs (U937, Daudi and Jurkat cells) were examined for their WT1, GATA-1, GATA-2 and Sp1 protein expressions by western blotting as described in Materials and methods. b-Actin was shown as the internal control. Right: solid tumor cell lines with high WT1 mRNA (TYK-nu-cPr, Az521, SW480) as well as the negative control of HepG2 cells were analyzed for their WT1, GATA-1 and GATA-2 protein levels.

Article Snippet: Western blotting of WT1, GATA-1, GATA-2 and Sp1 was carried out using the anti-WT1 antibody (clone 6F-H2, Dakocytomation, CA, USA), anti-GATA-1 (sc-265), anti-GATA-2 (sc-9008) and anti-Sp1 (sc-59) antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), respectively.

Techniques: Western Blot, Control, Negative Control

Journal: Leukemia

Article Title: GATA-1 and GATA-2 binding to 3' enhancer of WT1 gene is essential for its transcription in acute leukemia and solid tumor cell lines.

doi: 10.1038/leu.2009.13

Figure Lengend Snippet: Figure 3 Promoter analysis of the 50 region of WT1 gene. Using either K562 or Jurkat cells, the promoter activity of WT1 gene was examined by transfecting the pGL3 basic reporter vector containing various lengths of the 50 promoter region as described in Materials and methods (a). Results were shown as the ratio of luciferase activity/b-gal. Data of pGL3 basic without 50 promoter were regarded as 1.0. The Sp1 site located between 224 and 203 bp is illustrated in (b). The solid circle in (c) shows mutated Sp1 site. The mean±s.d. was calculated from three independent experiments.

Article Snippet: Western blotting of WT1, GATA-1, GATA-2 and Sp1 was carried out using the anti-WT1 antibody (clone 6F-H2, Dakocytomation, CA, USA), anti-GATA-1 (sc-265), anti-GATA-2 (sc-9008) and anti-Sp1 (sc-59) antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), respectively.

Techniques: Activity Assay, Plasmid Preparation, Luciferase

Journal: Leukemia

Article Title: GATA-1 and GATA-2 binding to 3' enhancer of WT1 gene is essential for its transcription in acute leukemia and solid tumor cell lines.

doi: 10.1038/leu.2009.13

Figure Lengend Snippet: Figure 4 Effects of the intron 3 and 30 enhancers of WT1 gene. Effects of the intron 3 and 30 enhancers were analyzed by luciferase reporter assay using various combinations of the promoter and enhancers as shown in (a) left. The sequence information was derived from earlier publications14,15,20 and is described in Supplementary information. The mean ±s.d. was calculated from three independent experiments. Data of the relative luciferase activity of control vector (without enhancer elements) of each cell line were regarded as 1.0. In (b and c) deletion and mutation were introduced as illustrated in the left part of each panel. The open circle denotes wild-type GATA site, whereas the solid circle shows mutated GATA site. Data of the relative luciferase activity from luc vector containing only 50 promoter and intron 3 enhancer were regarded as 1.0.

Article Snippet: Western blotting of WT1, GATA-1, GATA-2 and Sp1 was carried out using the anti-WT1 antibody (clone 6F-H2, Dakocytomation, CA, USA), anti-GATA-1 (sc-265), anti-GATA-2 (sc-9008) and anti-Sp1 (sc-59) antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), respectively.

Techniques: Luciferase, Reporter Assay, Sequencing, Derivative Assay, Activity Assay, Control, Plasmid Preparation, Mutagenesis

Journal: Leukemia

Article Title: GATA-1 and GATA-2 binding to 3' enhancer of WT1 gene is essential for its transcription in acute leukemia and solid tumor cell lines.

doi: 10.1038/leu.2009.13

Figure Lengend Snippet: Figure 5 Effect of modulation of GATA family genes. GATA-1 and GATA-2 expression vectors were transiently introduced to Jurkat and HepG2 cells whose original WT1 mRNA levels were low. Under overexpression conditions shown in the upper parts of a and b, we determined WT1 promoter/enhancer activities using the e2 reporter of Figure 4c (a), or WT1 protein levels (b). Promoter/enhancer activity was expressed with the mock transfectant regarded as 1.0. (c) siRNAs of GATA-1 and GATA-2 were transfected as described in the Materials and methods. At 48 h after transfection, the WT1 protein level was evaluated by western blotting with b-actin as the internal control ( right). Scramble siRNA was used as the control of the specificity of siRNAs used.

Article Snippet: Western blotting of WT1, GATA-1, GATA-2 and Sp1 was carried out using the anti-WT1 antibody (clone 6F-H2, Dakocytomation, CA, USA), anti-GATA-1 (sc-265), anti-GATA-2 (sc-9008) and anti-Sp1 (sc-59) antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), respectively.

Techniques: Expressing, Over Expression, Activity Assay, Transfection, Western Blot, Control

Journal: Pathology Oncology Research

Article Title: Papillary Endothelial Hyperplasia (Masson Tumor) of the Hand. Surgical and Pathological Consideration from Seven Cases Using New Vascular Markers

doi: 10.1007/s12253-020-00838-8

Figure Lengend Snippet: Summary of patient age, gender, location of lesion, appearance of the lesion, duration of symptoms , previous treatments, radiological examinations, surgical care, postoperative complications, recurrences, pathological markers

Article Snippet: Tissue sections of 3 μm thickness were cut from the blocks, followed by deparaffinization in xylene and retrieval using either the Bond Epitope Retrieval Solution 1 (pH ∼ 6) or the Bond Epitope Retrieval Solution 2 (pH ∼ 9) (Leica Microsystems, Wetzlar, Germany) at 99–100 °C for 20–30 min, and immunostained using a monoclonal mouse anti-ASMA antibody (clone:1A4; 1:400; Agilent DAKO, USA California Santa Clara), monoclonal mouse anti-human ERG (clone EPR 3864; ready to use; Ventana, Roche Diagnostics USA Indianapolis), monoclonal mouse anti-human WT1 (clone 6F- H2; 1:300; BioSB, USA Santa Barbara CA), monoclonal mouse anti-human CD31, (clone JC70A; 1:300; Agilent Dako, USA California Santa Clara) and polyclonal mouse anti-human Glut1 (1:100; Cell Marcque, USA Rocklin California).

Techniques: Biomarker Discovery, Functional Assay

Journal: Pathology Oncology Research

Article Title: Papillary Endothelial Hyperplasia (Masson Tumor) of the Hand. Surgical and Pathological Consideration from Seven Cases Using New Vascular Markers

doi: 10.1007/s12253-020-00838-8

Figure Lengend Snippet: Glut1 is negative in endothelial cells (arrows), only red blood cells are positive (arrowheads) ( a ). WT1 is clearly positive on endothelial cells ( b ), while ASMA displays pericytes ( c ). (Immunohistochemical stain with Glut1, WT1, and ASMA)

Article Snippet: Tissue sections of 3 μm thickness were cut from the blocks, followed by deparaffinization in xylene and retrieval using either the Bond Epitope Retrieval Solution 1 (pH ∼ 6) or the Bond Epitope Retrieval Solution 2 (pH ∼ 9) (Leica Microsystems, Wetzlar, Germany) at 99–100 °C for 20–30 min, and immunostained using a monoclonal mouse anti-ASMA antibody (clone:1A4; 1:400; Agilent DAKO, USA California Santa Clara), monoclonal mouse anti-human ERG (clone EPR 3864; ready to use; Ventana, Roche Diagnostics USA Indianapolis), monoclonal mouse anti-human WT1 (clone 6F- H2; 1:300; BioSB, USA Santa Barbara CA), monoclonal mouse anti-human CD31, (clone JC70A; 1:300; Agilent Dako, USA California Santa Clara) and polyclonal mouse anti-human Glut1 (1:100; Cell Marcque, USA Rocklin California).

Techniques: Immunohistochemical staining, Staining

Journal: Cell proliferation

Article Title: LRH-1 activation alleviates diabetes-induced podocyte injury by promoting GLS2-mediated glutaminolysis.

doi: 10.1111/cpr.13479

Figure Lengend Snippet: FIGURE 1 Activation of glutamine metabolism pathway in glomeruli from DKD mice. (A) Different groups of mice were sacrificed at 16 weeks from the onset of this study. (B) Urinary albumin-to-creatinine ratio (UACR) in different groups of mice. *p < 0.05, n = 6. (C) Representative transmission electron microscopy (TEM) pictures showing podocyte foot process effacement and glomerular basement membrane (GBM) thickness in mice with DKD. Scale bars: 2 μm. (D) Representative periodic acid–Schiff (PAS) staining, Terminal labelling (TUNEL) staining, and immunofluorescence staining (WT1-positive podocytes) in glomerulus from renal tissues. Scale bar: 25 μm. (E) Heatmap of the metabolite changes in glomeruli from different groups of mice. (F) Glomerular sclerosis index of the PAS staining, *p < 0.05 (n = 6). (G) Quantitative analyses of TUNEL-staining-positive cell per glomerulus in renal tissues. *p < 0.05 (n = 20). (H) Quantitative analyses of WT1-staining-positive podocytes per glomerulus in renal tissues. *p < 0.05 (n = 20). (I, J) The relative glomerular content of Gln and Glu. *p < 0.05 (n = 6).

Article Snippet: Rabbit anti-LRH-1 antibodies were obtained from Proteintech (Wuhan, China, 22460-1-AP); mouse anti-WT1 antibodies were purchased from Novus (Colorado, USA, NB110-60011); rabbit anti-GLS2 antibodies were purchased from Abcam (Cambridge, UK, ab113509), guinea pig anti-Synaptopodin antibody was obtained from Progen Biotechnik (Heidelberg, Germany, GP94); anti-GAPDH mouse monoclonal antibody, fluorescent secondary antibodies were purchased from Antgene (Wuhan, China, ANT324).

Techniques: Activation Assay, Transmission Assay, Electron Microscopy, Membrane, Staining, TUNEL Assay, Immunofluorescence

Journal: Cell proliferation

Article Title: LRH-1 activation alleviates diabetes-induced podocyte injury by promoting GLS2-mediated glutaminolysis.

doi: 10.1111/cpr.13479

Figure Lengend Snippet: FIGURE 2 Changes in critical enzymes of Gln catabolism in the glomerular podocytes of db/db mice. (A) Heatmap showing the changes in critical metabolic enzymes of glutaminolysis and amino acid catabolism. (B) Western blot assay showing expression of glomerular LRH-1 and GLS2. *p < 0.05 (n = 3). (C, D) Representative immunofluorescence staining of LRH-1 and GLS2 with WT1/Synaptopodin-positive podocytes in glomerulus from renal tissues among different groups. Scale bar: 25 μm. (E) Representative immunohistochemical staining images showing the reduction of LRH-1 and GLS2 in glomerulus from renal tissues of db/db mice. Scale bar: 25 μm. (F, G) Quantitative analyses of Western blot assay. *p < 0.05 (n = 3). (H) Schematic diagram of mitochondrial Gln metabolism.

Article Snippet: Rabbit anti-LRH-1 antibodies were obtained from Proteintech (Wuhan, China, 22460-1-AP); mouse anti-WT1 antibodies were purchased from Novus (Colorado, USA, NB110-60011); rabbit anti-GLS2 antibodies were purchased from Abcam (Cambridge, UK, ab113509), guinea pig anti-Synaptopodin antibody was obtained from Progen Biotechnik (Heidelberg, Germany, GP94); anti-GAPDH mouse monoclonal antibody, fluorescent secondary antibodies were purchased from Antgene (Wuhan, China, ANT324).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Immunohistochemical staining

Journal: Cell proliferation

Article Title: LRH-1 activation alleviates diabetes-induced podocyte injury by promoting GLS2-mediated glutaminolysis.

doi: 10.1111/cpr.13479

Figure Lengend Snippet: FIGURE 5 DLPC upregulated LRH-1 and GLS2 expression. (A) Mice were treated daily by oral gavage with DLPC (100 mg/kg) or the solvent vehicle for 3 weeks and were sacrificed at 16 weeks after the onset of diabetes. (B) Western blot assay showing expression of LRH-1 and GLS2 among different groups. (C, D) Quantitative analyses of Western blot assay. *p < 0.05 (n = 3). (E, F) Representative immunofluorescence staining of LRH-1 and GLS2 with WT1/Synaptopodin-positive podocytes in glomerulus from renal tissues among different groups. Scale bar: 25 μm.

Article Snippet: Rabbit anti-LRH-1 antibodies were obtained from Proteintech (Wuhan, China, 22460-1-AP); mouse anti-WT1 antibodies were purchased from Novus (Colorado, USA, NB110-60011); rabbit anti-GLS2 antibodies were purchased from Abcam (Cambridge, UK, ab113509), guinea pig anti-Synaptopodin antibody was obtained from Progen Biotechnik (Heidelberg, Germany, GP94); anti-GAPDH mouse monoclonal antibody, fluorescent secondary antibodies were purchased from Antgene (Wuhan, China, ANT324).

Techniques: Expressing, Solvent, Western Blot, Immunofluorescence, Staining

Journal: Cell proliferation

Article Title: LRH-1 activation alleviates diabetes-induced podocyte injury by promoting GLS2-mediated glutaminolysis.

doi: 10.1111/cpr.13479

Figure Lengend Snippet: FIGURE 6 LRH-1 activation attenuates podocyte injury and renal injury in DKD mice. (A) Representative periodic acid–Schiff (PAS) staining, Terminal labeling (TUNEL), and immunofluorescence staining (WT1-positive podocytes) in glomerulus from renal tissues. Scale bar: 25 μm. (B) Representative transmission electron microscopy (TEM) pictures of glomerulus among different groups. Scale bars: 2 μm. (C) Urinary albumin- to-creatinine ratio (UACR) in different groups of mice. *p < 0.05, n = 6. (D) Glomerular sclerosis index of the PAS staining, *p < 0.05 (n = 6). (E) Quantitative analyses of TUNEL-staining-positive cell per glomerulus in renal tissues. *p < 0.05 (n = 20). (F) Quantitative analyses of WT1-staining-positive podocytes per glomerulus in renal tissues. *p < 0.05 (n = 20). (G, H) The relative glomerular content of Gln and Glu. *p < 0.05 (n = 6).

Article Snippet: Rabbit anti-LRH-1 antibodies were obtained from Proteintech (Wuhan, China, 22460-1-AP); mouse anti-WT1 antibodies were purchased from Novus (Colorado, USA, NB110-60011); rabbit anti-GLS2 antibodies were purchased from Abcam (Cambridge, UK, ab113509), guinea pig anti-Synaptopodin antibody was obtained from Progen Biotechnik (Heidelberg, Germany, GP94); anti-GAPDH mouse monoclonal antibody, fluorescent secondary antibodies were purchased from Antgene (Wuhan, China, ANT324).

Techniques: Activation Assay, Staining, Labeling, TUNEL Assay, Immunofluorescence, Transmission Assay, Electron Microscopy