Journal: STAR Protocols

Article Title: Protocol for potent activation of T cells using BPI-stimulated murine bone marrow-derived cells

doi: 10.1016/j.xpro.2026.104519

Figure Lengend Snippet: Expected results of this protocol Naïve CD4 + T cells cultured for d5 in supernatant of untreated BMDCs (SN NT) or in supernatant of BPI-treated BMDCs (SN BPI). (A) Representative dot blot of flow cytometric analysis of CD62L and CD44 cell surface presentation. (B) IL-22 secretion measured by Luminex technology, n = 4. Data are shown as means ± SEM. Statistical testing was performed using Student`s ratio paired t test.

Article Snippet: CD44 Antibody, anti-mouse, PE-Vio 770 clone IM7.8.1; 1:10 dilution , Miltenyi Biotec , Cat# 130-102-377 RRID: AB_2658187.

Techniques: Cell Culture, Dot Blot, Luminex

Journal: Molecular Therapy Oncology

Article Title: Chimeric antigen receptor T cell targeting CD44E variant in HCC holds therapeutic potential

doi: 10.1016/j.omton.2026.201181

Figure Lengend Snippet: Live-cell fluorescence microscopy capturing the behavior of mock CAR T cells (white) co-cultured with HKCI-C1 CD44E⁺ cells (green) over 24 h, serving as a negative control.

Article Snippet: For CD44E expression, TaqMan assay for CD44E (Hs01081475_m1) and internal control 18S rRNA (Hs99999901_s1) were purchased from Applied Biosystems.

Techniques:

Journal: Molecular Therapy Oncology

Article Title: Chimeric antigen receptor T cell targeting CD44E variant in HCC holds therapeutic potential

doi: 10.1016/j.omton.2026.201181

Figure Lengend Snippet: Time-lapse fluorescence microscopy capturing the interaction and killing kinetics of anti-CD44E CAR T cells (white) co-cultured with HKCI-C1 CD44E⁺ cells (green) over 24 h.

Article Snippet: For CD44E expression, TaqMan assay for CD44E (Hs01081475_m1) and internal control 18S rRNA (Hs99999901_s1) were purchased from Applied Biosystems.

Techniques:

Journal: Molecular Therapy Oncology

Article Title: Chimeric antigen receptor T cell targeting CD44E variant in HCC holds therapeutic potential

doi: 10.1016/j.omton.2026.201181

Figure Lengend Snippet: Real-time dynamics of anti-CD44E CAR T cells (red) targeting and eliminating 3D liver cancer organoid Org_688T cells (green) during a 24-h co-culture period.

Article Snippet: For CD44E expression, TaqMan assay for CD44E (Hs01081475_m1) and internal control 18S rRNA (Hs99999901_s1) were purchased from Applied Biosystems.

Techniques:

Journal: Molecular Therapy Oncology

Article Title: Chimeric antigen receptor T cell targeting CD44E variant in HCC holds therapeutic potential

doi: 10.1016/j.omton.2026.201181

Figure Lengend Snippet: CD44E-specific antibody screening and selection (A) The titers of anti-CD44E antibodies produced by hybridoma colonies were determined by ELISA. Anti-pan CD44 antibodies were selected as controls. (B) Binding of anti-CD44E-positive hybridoma supernatants to CD44E + cells was analyzed by flow cytometry. (C) Binding of anti-pan CD44-positive hybridoma supernatants to CD44E + cells was analyzed by flow cytometry. (D) Serial dilutions of purified anti-CD44E antibody (from S431 hybridoma colony) were added to ELISA plates coated with hCD44E-Fc and hCD44-Fc proteins to measure their binding activity and determine EC 50 values. (E) Surface plasmon resonance (SPR) analysis of S431 antibody binding to human CD44 isoforms. Real-time binding sensorgrams showing the interaction of serially diluted human CD44E-hFc (left) and human CD44-hFc (right) with immobilized S431 antibody. The binding response is indicated in resonance units (RU). (F) Flow cytometric analysis of antibody-binding specificity. The binding of the anti-CD44E antibody (S431 clone) was evaluated in HKCI-C1 cells overexpressing CD44E or CD44S (left), while the binding of anti-pan CD44 antibody (S129 clone) was assessed in the same cells (right). Human IgG (hIgG) was used as an isotype control. (G) Immunohistochemical (IHC) staining of CD44E in primary human liver tumor tissues (2 representative cases). Scale bars, 200 μm. (H) Representative immunofluorescent staining detecting cell-surface expression of CD44E protein in HKCI-11 cells. Scale bars, 100 μm.

Article Snippet: For CD44E expression, TaqMan assay for CD44E (Hs01081475_m1) and internal control 18S rRNA (Hs99999901_s1) were purchased from Applied Biosystems.

Techniques: Selection, Produced, Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Purification, Activity Assay, SPR Assay, Control, Immunohistochemical staining, Immunohistochemistry, Staining, Expressing