Journal: Oncology Letters

Article Title: Solitary fibrous tumor of the brain: A report of 3 cases

doi: 10.3892/ol.2024.14621

Figure Lengend Snippet: Clinicopathological parameters of the reported cases.

Article Snippet: Immunostaining for STAT6 (rabbit monoclonal antibody clone EP325; 1:100; cat. no. BSB-3809-05; Bio SB Inc.), CD34 (rabbit monoclonal antibody clone EP88; 1:100; cat. no. 134R-16-RUO; Cell Marque; Merck KGaA) and Ki67 (mouse monoclonal antibody clone MIB-1; 1:100; cat. no. Z2305ML; Zeta Corporation) at 37°C for 44 min was conducted using a Ventana BenchMark XT 750-700 Automated IHC/ISH autostainer (Roche Diagnostics, Ltd.) was performed as per the manufacturer's protocol, using heat-induced antigen retrieval (100°C, 4 min) and endogenous peroxidase blocking by 0.04% hydrogen peroxide.

Techniques: Imaging, Immunohistochemistry, Biomarker Discovery

Journal: Oncology Letters

Article Title: Solitary fibrous tumor of the brain: A report of 3 cases

doi: 10.3892/ol.2024.14621

Figure Lengend Snippet: Microscopic images of Case 1. (A) H&E-stained image (magnification, ×10). Tumor cells were observed in sheets with interspersed dilated vessels (*). (B) H&E-stained image (magnification, ×20). The arrow highlights the amianthoid fibers. (C) H&E-stained images (magnification, ×40). The arrow highlights the mitotic figures. (D) H&E-stained image (magnification, 10×), *indicates necrosis. Immunohistochemistry (DAB stain; magnification, ×20) for (E) CD34 and (F) STAT6 was positive. H&E, hematoxylin and eosin.

Article Snippet: Immunostaining for STAT6 (rabbit monoclonal antibody clone EP325; 1:100; cat. no. BSB-3809-05; Bio SB Inc.), CD34 (rabbit monoclonal antibody clone EP88; 1:100; cat. no. 134R-16-RUO; Cell Marque; Merck KGaA) and Ki67 (mouse monoclonal antibody clone MIB-1; 1:100; cat. no. Z2305ML; Zeta Corporation) at 37°C for 44 min was conducted using a Ventana BenchMark XT 750-700 Automated IHC/ISH autostainer (Roche Diagnostics, Ltd.) was performed as per the manufacturer's protocol, using heat-induced antigen retrieval (100°C, 4 min) and endogenous peroxidase blocking by 0.04% hydrogen peroxide.

Techniques: Staining, Immunohistochemistry

Journal: Oncology Letters

Article Title: Solitary fibrous tumor of the brain: A report of 3 cases

doi: 10.3892/ol.2024.14621

Figure Lengend Snippet: Microscopic images of Case 2. (A) H&E-stained image (magnification, ×10). ‘Patternless’ architecture of tumor cells with a hemangiopericytomatous vasculature (*). (B) H&E-stained image (magnification, ×40). The arrow highlights mitotic figures. Immunohistochemistry (DAB stain; magnification, ×10) for (C) CD34 was negative, while (D) STAT6 was positive in the tumor cells. H&E, hematoxylin and eosin.

Article Snippet: Immunostaining for STAT6 (rabbit monoclonal antibody clone EP325; 1:100; cat. no. BSB-3809-05; Bio SB Inc.), CD34 (rabbit monoclonal antibody clone EP88; 1:100; cat. no. 134R-16-RUO; Cell Marque; Merck KGaA) and Ki67 (mouse monoclonal antibody clone MIB-1; 1:100; cat. no. Z2305ML; Zeta Corporation) at 37°C for 44 min was conducted using a Ventana BenchMark XT 750-700 Automated IHC/ISH autostainer (Roche Diagnostics, Ltd.) was performed as per the manufacturer's protocol, using heat-induced antigen retrieval (100°C, 4 min) and endogenous peroxidase blocking by 0.04% hydrogen peroxide.

Techniques: Staining, Immunohistochemistry

Journal: Oncology Letters

Article Title: Solitary fibrous tumor of the brain: A report of 3 cases

doi: 10.3892/ol.2024.14621

Figure Lengend Snippet: Microscopic images of subpleural metastatic nodule in Case 3. (A) H&E-stained image (magnification, ×10). Low-power magnification of fibromuscular tissue infiltrated by tumor cells. (B) H&E-stained image (magnification, ×20). *marks the area with stromal hyalinization. Immunohistochemistry (DAB stain; magnification, ×20) for (C) CD34 shows heterogenous positivity, while (D) STAT6 shows diffuse positive expression. H&E, hematoxylin and eosin.

Article Snippet: Immunostaining for STAT6 (rabbit monoclonal antibody clone EP325; 1:100; cat. no. BSB-3809-05; Bio SB Inc.), CD34 (rabbit monoclonal antibody clone EP88; 1:100; cat. no. 134R-16-RUO; Cell Marque; Merck KGaA) and Ki67 (mouse monoclonal antibody clone MIB-1; 1:100; cat. no. Z2305ML; Zeta Corporation) at 37°C for 44 min was conducted using a Ventana BenchMark XT 750-700 Automated IHC/ISH autostainer (Roche Diagnostics, Ltd.) was performed as per the manufacturer's protocol, using heat-induced antigen retrieval (100°C, 4 min) and endogenous peroxidase blocking by 0.04% hydrogen peroxide.

Techniques: Staining, Immunohistochemistry, Expressing

Journal: International Journal of Biological Sciences

Article Title: USP25 stabilizes STAT6 to promote IL-4-induced macrophage M2 polarization and fibrosis

doi: 10.7150/ijbs.99345

Figure Lengend Snippet: USP25 inhibit K48 ubiquitination of STAT6 to enhance STAT6/PPAR-γ signaling in macrophages. (A) Upper panel: representative Western blot results for STAT6, PPAR-γ and SOCS3 at different time points stimulated with IL-4. Lower panel: figures showing the data with three mice analyzed. (B) Western blot analysis of SOCS3 in the lung homogenates. (C) USP25 did not affect MAPK (p38 and ERK), Akt and PI3K signaling. (D) Upper panel: immunoprecipitation of proteins using USP25 antibody and immunoblotting analysis using STAT6 antibody. Lower panel: immunoprecipitation of proteins using STAT6 antibody and immunoblotting analysis using USP25 antibody. (E) BMDMs of WT and USP25 -/- were treated with MG132 (10 μM) for 6 h before lysis, followed by stimulation of IL-4 (10 ng/mL) for 30 min. Proteins were immunoprecipitated with STAT6 antibody and analyzed by immunoblotting with the indicated antibodies. (F) NIH/3T3 cells were transfected with plasmids encoding the indicated constructs. Proteins were immunoprecipitated with anti-STAT6 antibody and analyzed by immunoblotting with the indicated antibodies. IP, immunoprecipitation; STAT6, signal transducer and activator of transcription 6; PPAR-γ, peroxisome proliferator-activated receptor gamma; SOCS3, suppressor of cytokine signaling 3. *p < 0.05 and **p < 0.01.

Article Snippet: Primary antibodies include anti-STAT6 antibody (Cat:380957, ZENBIO, China), anti PPAR-γ antibody (Cat: A11183, ABCLONAL, China), anti-USP25 antibody (Cat: ab187156, Abcam, USA), anti-Arg1 antibody (Cat: ab91279, Abcam, USA), anti-Ubi antibody (Cat: 134953, Abcam, USA), anti-Ub-k48 antibody (Cat: 140601, Abcam, USA).

Techniques: Ubiquitin Proteomics, Western Blot, Immunoprecipitation, Lysis, Transfection, Construct

Journal: Immunity

Article Title: Complex Autoinflammatory Syndrome Unveils Fundamental Principles of JAK1 Kinase Transcriptional and Biochemical Function

doi: 10.1016/j.immuni.2020.07.006

Figure Lengend Snippet:

Article Snippet: anti-pSTAT6 149 Sm-conjugated Clone 18 , Fluidigm , Cat No.3149004A.

Techniques: Recombinant, Virus, Molecular Cloning, Blocking Assay, Conjugation Assay, Staining, Western Blot, Lysis, Extraction, Mutagenesis, Isolation, Reverse Transcription, Luminex, Transfection, Amplification, Software, Variant Assay

Journal: Cell Death and Differentiation

Article Title: PP2Acα promotes macrophage accumulation and activation to exacerbate tubular cell death and kidney fibrosis through activating Rap1 and TNFα production

doi: 10.1038/s41418-021-00780-5

Figure Lengend Snippet: a , b Western blot assay ( a ) and semi-quantitative analysis ( b ) showing that deletion of PP2Acα enhanced LPS-stimulated Stat6 phosphorylation in BMDMs. * p < 0.05, n = 3. Data are presented as means ± SEM. c , d Western blot assay ( c ) and quantitative analysis ( d ) showing that overexpression of PP2Acα could inhibit Stat6 phosphorylation caused by PP2Acα ablation. * p < 0.05, n = 3. # p < 0.05, n = 3. Data are presented as means ± SEM. e Real-time qRT-PCR analysis showing the mRNA abundance of Tnfα in BMDMs. * p < 0.05, n = 6. # p < 0.05, n = 6. Data are presented as means ± SEM. f ELISA assay showing the protein abundance of TNFα in the cultural media of AS1517499-treated BMDMs. * p < 0.05, n = 3. # p < 0.05, n = 3. $ p < 0.05, n = 3. Data are presented as means ± SEM. g , h Representative PI staining images ( g ) and quantitative analysis ( h ) showing that the conditioned media of AS1517499-treated macrophages caused more tubular cell death. Scale bar, 100 μm. * p < 0.05, n = 3. # p < 0.05, n = 3. $ p < 0.05, n = 3. Data are presented as means ± SEM. i Real-time qRT-PCR analysis showing the mRNA abundance of Stat6 in scramble siRNA and Stat6 siRNA-transfected BMDMs. * p < 0.05, n = 3. Data are presented as means ± SEM. j ELISA analysis showing the protein abundance of TNFα in the cultural media of Stat6 siRNA-transfected BMDMs. * p < 0.05, n = 3. # p < 0.05, n = 3. $ p < 0.05, n = 3. Data are presented as means ± SEM. k , l Representative PI staining images ( k ) and quantitative analysis ( l ) showing that the conditioned media of Stat6 siRNA-transfected macrophages caused more tubular cell death. Scale bar, 100 μm. * p < 0.05, n = 3. # p < 0.05, n = 3. $ p < 0.05, n = 3. Data are presented as means ± SEM. m Representative immune staining images showing the induction of p-Stat6 (T645) in F4/80-positive macrophages within MФ-PP2Acα −/− fibrotic kidneys. White arrows indicate double-staining-positive cells. Scale bar, 10 μm. n , o Quantitative analysis for p-Stat6 (T645) and F4/80 double-positive cells in IRI ( n ) and UUO ( o ) kidney tissues. * p < 0.05, n = 3. Data are presented as means ± SEM. p , q Western blot analyses ( p ) and quantitative determination ( q ) showing that deletion of PP2Acα could induce the phosphorylation of Stat6 in macrophages sorted from MФ-PP2Acα −/− IRI kidneys. * p < 0.05, n = 3. Data are presented as means ± SEM. r , s Western blot analyses ( r ) and quantitative determination ( s ) showing that deletion of PP2Acα could induce the phosphorylation of Stat6 in macrophages sorted from MФ-PP2Acα −/− UUO kidneys. * p < 0.05, n = 3. Data are presented as means ± SEM.

Article Snippet: The primary antibodies were anti-PP2Ac (cat: 2038, Cell Signaling Technology, Boston, MA, USA, 1:1000), anti-PP2Acα (cat: ab106262, Abcam, Cambridge, UK, 1:1000), anti-PP2Acβ (cat: ab168371, Abcam, 1:1000), anti-methyl-PP2Ac (L309) (cat: ab66597, Abcam, 1:1000), anti-Itgb2 (cat: ab119830, Abcam, 1:1000), anti-Epac1 (cat: ab124162, Abcam, 1:1000), anti-FN (cat: F3648, Sigma-Aldrich, 1:10000), anti-Stat6 (cat: ab32520, Abcam, 1:1000), anti-p-Stat6 (T645) (cat: BS4186, Bioworld Technology, Nanjing, China, 1:1000), anti-Rap1a/b (cat: 4938, Cell Signaling Technology, 1:1000), anti-tubulin (cat: sc53646, Santa Cruz Biotechnology, 1:10000), and anti-GAPDH (cat: FL-335, Santa Cruz Biotechnology, 1:5000).

Techniques: Western Blot, Over Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Transfection, Double Staining