Journal: Cell Reports Medicine

Article Title: Synergistic T cell signaling by 41BB and CD28 is optimally achieved by membrane proximal positioning within parallel chimeric antigen receptors

doi: 10.1016/j.xcrm.2021.100457

Figure Lengend Snippet:

Article Snippet: Anti-human CD98 - PE-Vio770 , Miltenyi Biotec , Cat# 130-105-710, RRID: AB_2659686.

Techniques: Control, Recombinant, Staining, Transfection, Enzyme-linked Immunosorbent Assay, Luciferase, Gene Expression, Cloning, Software

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: A negative feedback loop between fibroadipogenic progenitors and muscle fibres involving endothelin promotes human muscle fibrosis

doi: 10.1002/jcsm.12974

Figure Lengend Snippet: Antibody panel used for CyTOF

Article Snippet: CD98 , 159 Tb , UM7F8 , Fluidigm , 3159022B.

Techniques:

Journal: Materials Today Bio

Article Title: A bioengineered tumor matrix-based scaffold for the evaluation of melatonin efficacy on head and neck squamous cancer stem cells

doi: 10.1016/j.mtbio.2024.101246

Figure Lengend Snippet: HNSCC microenvironment model. (A) Characterization of Cal-27 CSCs. Image of tumorspheres grown in suspension with serum-free spheres medium (10×). Cytometry histograms of CD98 and CD44 CSCs cell-membrane markers expression. Percentage of ALDH1 activity of cells grown in monolayer or in suspension (cytometry histograms besides). (B) Schematic representation of the TME components embedded in the hydrogel. Confocal representative images of cell viability of tumorspheres, FBs and MSCs (stroma) and the co-culture of stroma and tumorspheres (TME) cultured in the hydrogel. Living cells shown in green and nuclei of death cells in red. Scale bar: 200 μm. (C) Proliferation assay of the tumorspheres, FBs and MSCs (stroma) and the co-culture of stroma and tumorspheres (TME) cultured in the hydrogel. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Cells were centrifuged followed by the addition of fluorochrome-conjugated monoclonal antibodies for CD98 and CD44 (Miltenyi Biotec) according to the manufacturer's instructions and incubated at 4 °C in the dark for 12 min. After adding BSA, cells were centrifugated and resuspended in PBS and analyzed by flow cytometry in a FACSCanto II cytometer (BD Biosciences).

Techniques: Suspension, Cytometry, Membrane, Expressing, Activity Assay, Co-Culture Assay, Cell Culture, Proliferation Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A monoclonal antibody recognizing CD98 on human embryonic stem cells shows anti-tumor activity in hepatocellular carcinoma xenografts

doi: 10.1007/s00262-024-03827-x

Figure Lengend Snippet: Flow cytometric analysis of various cells with NPB15. a Binding profiles of NPB15 were examined by flow cytometry in naïve and primed hESCs, mouse embryonic fibroblasts (MEFs) and retinoic acid-treated primed H9 cells. b Binding profiles of NPB15 were examined by flow cytometry in various cancer cell lines and PBMCs and hepatocytes

Article Snippet: Primary antibodies used were NPB15, Chi-NPB15, anti-human CD98 (Ancell corporation, Bayport, MN, USA), anti-mouse CD98 (BioLegend, San Diego, CA, USA), anti-CD133 (Abcam, Cambridge, UK), anti-EpCAM (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-CD44 (Invitrogen, Waltham, MA, USA).

Techniques: Binding Assay, Flow Cytometry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A monoclonal antibody recognizing CD98 on human embryonic stem cells shows anti-tumor activity in hepatocellular carcinoma xenografts

doi: 10.1007/s00262-024-03827-x

Figure Lengend Snippet: Identification and validation of cell surface molecule recognized by NPB15. a A549 cell lysates were immunoprecipitated with NPB15 after cell surface biotinylation, and the immunoprecipitants were detected with SA-HRP in Western blotting. Preclearing was done with protein G-agarose beads alone and used as a control (Con). b A549 cell lysates were immunoprecipitated with NPB15, and the immunoprecipitants were detected with α -CD98 in Western blotting. c , d Immunoprecipitation of FLA-tagged CD98 with mouse IgG (mIgG), NPB15, α -CD98 and α -FLAG antibodies after overexpression of FLAG-tagged CD98 in HEK293FT cells. Immunoprecipitants were analyzed by Western blotting with α -CD98 ( c ) or α -FLAG antibodies ( d ). Preclearing was done with protein G-agarose beads alone and used as a control

Article Snippet: Primary antibodies used were NPB15, Chi-NPB15, anti-human CD98 (Ancell corporation, Bayport, MN, USA), anti-mouse CD98 (BioLegend, San Diego, CA, USA), anti-CD133 (Abcam, Cambridge, UK), anti-EpCAM (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-CD44 (Invitrogen, Waltham, MA, USA).

Techniques: Immunoprecipitation, Western Blot, Control, Over Expression

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A monoclonal antibody recognizing CD98 on human embryonic stem cells shows anti-tumor activity in hepatocellular carcinoma xenografts

doi: 10.1007/s00262-024-03827-x

Figure Lengend Snippet: CD98 depletion inhibits cell proliferation by promoting apoptosis in HCC cells. a CD98 protein expression of the control (siCon) and CD98 knockdown (siCD98) SNU449 and Huh7 cells. Indicated protein levels were analyzed by Western blotting and Image J software. b Flow cytometry of the CD98 expression in siCon and siCD98 SNU449 and Huh7 using NPB15. c Quantification of cell numbers of siCon and siCD98 SNU449 and Huh7 cells (* p < .05; *** p < .005). d, e Apoptosis analysis of siCon and siCD98 SNU449 and Huh7 cells. Cells were stained with PI and annexin V. f, g Quantification of d and e (ns, not significant; ** p < .01; *** p < .005)

Article Snippet: Primary antibodies used were NPB15, Chi-NPB15, anti-human CD98 (Ancell corporation, Bayport, MN, USA), anti-mouse CD98 (BioLegend, San Diego, CA, USA), anti-CD133 (Abcam, Cambridge, UK), anti-EpCAM (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-CD44 (Invitrogen, Waltham, MA, USA).

Techniques: Expressing, Control, Knockdown, Western Blot, Software, Flow Cytometry, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A monoclonal antibody recognizing CD98 on human embryonic stem cells shows anti-tumor activity in hepatocellular carcinoma xenografts

doi: 10.1007/s00262-024-03827-x

Figure Lengend Snippet: CD98 expression is positively associated with cancer stemness. a Cell surface expression of CD44 was examined in CD98 knockdown SNU449 cells by flow cytometry. b Quantification of a . The MFIs of NPB15 and CD44 were quantified and normalized against the MFIs of control cells (siCon). c Protein expression of EpCAM and CD13 was analyzed in CD98 knockdown (siCD98) Huh7 cells by Western blotting. d Quantification of c . Protein expression levels of siCD98 Huh7 cells were quantified by Image J software and normalized against control cells (siCon). α-tubulin was used as a loading control. e ALDH activity of CD98 knockdown cancer cells. ALDH activity was measured and normalized against control cells (siCon). f Image of tumor spheroids (scale bar, 64.3 µm). g, h Flow cytometry analysis of Huh7 tumor spheroids with CSC markers. The expression levels of CD133, EpCAM, CD44 and CD98 were quantified and normalized against those of adherent cells (* p < .05; ** p < .01)

Article Snippet: Primary antibodies used were NPB15, Chi-NPB15, anti-human CD98 (Ancell corporation, Bayport, MN, USA), anti-mouse CD98 (BioLegend, San Diego, CA, USA), anti-CD133 (Abcam, Cambridge, UK), anti-EpCAM (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-CD44 (Invitrogen, Waltham, MA, USA).

Techniques: Expressing, Knockdown, Flow Cytometry, Control, Western Blot, Software, Activity Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A monoclonal antibody recognizing CD98 on human embryonic stem cells shows anti-tumor activity in hepatocellular carcinoma xenografts

doi: 10.1007/s00262-024-03827-x

Figure Lengend Snippet: NPB15-sorted CD98-high cells show higher clonogenic survival than CD98-low cells. a SNU449 cells were stained with NPB15, and the top 30% (CD98-high) and bottom 30% cell populations (CD98-low) were sorted on a cytometer. b NPB15-Sorted CD98-high and -low cells were subjected to clonogenic survival assays. The scale bar is 10 mm. c Quantification of b. The number of colonies was quantified by Image J software (*** p < .005)

Article Snippet: Primary antibodies used were NPB15, Chi-NPB15, anti-human CD98 (Ancell corporation, Bayport, MN, USA), anti-mouse CD98 (BioLegend, San Diego, CA, USA), anti-CD133 (Abcam, Cambridge, UK), anti-EpCAM (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-CD44 (Invitrogen, Waltham, MA, USA).

Techniques: Staining, Cytometry, Software

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: A monoclonal antibody recognizing CD98 on human embryonic stem cells shows anti-tumor activity in hepatocellular carcinoma xenografts

doi: 10.1007/s00262-024-03827-x

Figure Lengend Snippet: NPB15 shows antitumor activity in an HCC xenograft animal model. a ADCC assay of chimeric NPB15 antibody on Huh7 cells. Huh7 cells were treated with 0 to 100 μg/ml Chi-NPB15 or human IgG1 isotype control and incubated with effector Jurkat cell for 6 h. The relative luminescence unit (RLU) is plotted against the logarithmic antibody concentration (* p < .05; *** p < .005). b A schematic diagram showing the workflow of antibody injection into Huh7 xenograft mice. BALB/c nu/nu mice were injected subcutaneously with Huh7 cells and injected intravenously every 2 days with mIgG1 or NPB15 after the tumor volume reached 100 mm 3 . c Graphical representation of mIGg1- and NPB15-treated tumor growth, respectively. Tumor width and length were determined using a caliper. Data are presented as mean values ± SEM for isotype ( n = 6) and NPB15 ( n = 6) (* p < .05; *** p < .005). d Tumors from killed xenograft models are shown. e Tumors extracted (shown in d ) were weighted and mean tumor weights ± SEM are presented (* p < 0.05). f Effects of antibody injection on body weight of HCC xenograft mice. Body weights of 12 mice were measured every 2 days and mean body weights ± SEM are presented (ns, not significant). g Western blot analysis of CD98 in NPB15- or mIgG1-treated tumor tissues. Formaldehyde-treated tumor tissues were homogenized and analyzed by Western blot analysis with anti-CD98 antibody. α -tubulin was used as an internal loading control. CL CD98, cross-linked CD98 by HCHO

Article Snippet: Primary antibodies used were NPB15, Chi-NPB15, anti-human CD98 (Ancell corporation, Bayport, MN, USA), anti-mouse CD98 (BioLegend, San Diego, CA, USA), anti-CD133 (Abcam, Cambridge, UK), anti-EpCAM (Santa Cruz Biotechnology, Dallas, TX, USA) and anti-CD44 (Invitrogen, Waltham, MA, USA).

Techniques: Activity Assay, Animal Model, ADCC Assay, Control, Incubation, Concentration Assay, Injection, Western Blot