Journal: Cancer Science

Article Title: Radioimmunotherapy of human synovial sarcoma using a monoclonal antibody against FZD10

doi: 10.1111/j.1349-7006.2007.00701.x

Figure Lengend Snippet: Immunohistochemical staining analyses of synovial sarcoma and normal human tissue sections using MAb 92‐13; (a,b) synovial sarcoma tissue sections (c) colon, (d) heart, (e) lung, (f) liver, (g) kidney and (h) placenta. Original magnification ×100. Bar, 50 µm.

Article Snippet: For immunohistochemical staining analysis, 4‐µm sections of paraffin‐embedded normal adult human tissues (lung, heart, liver, kidney, colon and placenta) (BioChain, Hayward, CA, USA) and surgical synovial sarcoma specimens were deparaffinized and processed for antigen retrieval in Target Retrieval Solution (pH 9) (DAKO Cytomation, Carpinteria, CA, USA) at 125°C for 30 s. After treatment with peroxidase blocking reagent (DAKO Cytomation) for 10 min, followed by treatment with protein blocking serum‐free (DAKO Cytomation) for 30 min, slides were incubated with 10 µg/mL MAb 92‐13.

Techniques: Immunohistochemical staining, Staining

Journal: International Journal of Oncology

Article Title: Long non-coding RNA nuclear paraspeckle assembly transcript 1 regulates ionizing radiation-induced pyroptosis via microRNA-448/gasdermin E in colorectal cancer cells

doi: 10.3892/ijo.2021.5259

Figure Lengend Snippet: lncRNA NEAT1 upregulates GSDME via miR-448. (A) GSDME expression in human COAD was obtained from Human Protein Atlas ( http://www.proteinatlas.org ). The examples of the pictures by sex and patient id are shown and the images are available at v.20.1 www.proteinatlas.org . (B) Expression of lncRNA NEAT1 and GSDME in the major stages of COAD. (C) Overall survival analysis of NEAT1 and GSDME in COAD from the Cancer Genome Atlas database using the Gene Expression Profiling Interactive Analysis online tool. (D-G) HCT116 cells were transfected with siGSDME, siNEAT1-1, siNEAT1-2, siNEAT1-1+2, or negative control siRNA. After 48 h, (D) NEAT1 expression and (E) GSDME expression were analyzed using RT-qPCR. GAPDH was used as a control. (F) miR-448 expression was analyzed using RT-qPCR in HCT116 cells after transfection with siNEAT1-1, siNEAT1-2 and siNEAT1-1+2. (G) Expression of full-length and cleaved GSDME was analyzed using western blotting. GAPDH was used as a loading control. (H) GSDME mRNA expression was analyzed using RT-qPCR in HCT116 cells after transfection with siGSDME, siNEAT1-1+2, or co-transfection with siNEAT1-1+2 and miR-448 mimetics. * P<0.05, ** P<0.01 compared with the NC group unless otherwise indicated. RT-q, reverse transcription-quantitative; GSDME, gasdermin E; miR, microRNA; si, small interfering; NEAT1, nuclear paraspeckle assembly transcript 1; lncRNA, long non-coding RNA; COAD, colon adenocarcinoma; HR, hazard ratio; NC, negative control (comprising cells transfected with NC siRNA); normal, untransfected cells.

Article Snippet: Immunostaining images of GSDME in normal colon and colon adenocarcinoma tissues were obtained from the Human Protein Atlas ( http://www.proteinatlas.org ).

Techniques: Expressing, Gene Expression, Transfection, Negative Control, Quantitative RT-PCR, Control, Western Blot, Cotransfection, Reverse Transcription

Journal: Toxins

Article Title: Shiga Toxin Is Transported into the Nucleoli of Intestinal Epithelial Cells via a Carrier-Dependent Process

doi: 10.3390/toxins2061318

Figure Lengend Snippet: Localization of Stx and StxB to the nucleoli of intestinal epithelial T84 cells and human colonic tissue. ( a ) Stx-Alexa 488 (green) appears in the nucleoli of PM-permeabilized confluent T84 cells; blue: nuclear staining by Hoechst dye; arrows point to the nucleoli filled with Stx-Alexa 488. ( b ) Recombinant StxB-Alexa 568 (red) is delivered from cytoplasm into the nucleoli of PM-permeabilized confluent T84 cells; blue: nuclear staining by Hoechst dye. ( c ) T84 cells immunostained with an antibody against the nucleolar protein nucleolin (red). ( d ) Same cells as in c stained with StxB-Alexa 488 (green), confirming StxB nucleolar localization. ( e ) Normal human colonic tissue immunostained with StxB-Alexa 488 (green) indicates the presence of StxB binding sites in nucleoli of crypt epithelial cells; red: β-catenin; blue: Hoechst nuclear stain; arrows in the magnified insert point to the nucleoli inside the nuclei filled with StxB-Alexa 488. ( f ) StxB-Alexa 568 (red) is accumulated in the nucleoli of PM-permeabilized T84 cells, while CTB-fluorescein (green) is present in cytoplasm and membranes but not in nucleoli.

Article Snippet: Human normal colonic tissue: Frozen 20 μm thick samples of surgically removed, normal human colonic tissue were obtained from the Johns Hopkins Colorectal Cancer Registry with JHM-IRB1 granted exemption.

Techniques: Staining, Recombinant, Binding Assay