mouse monoclonal anti osteocalcin antibodies  (R&D Systems)

Journal: bioRxiv

Article Title: Ablation of glypican-3 enhances radiosensitivity in liver cancer by prolonging G2/M arrest and activating the ATM/CHK2 pathway

doi: 10.64898/2026.05.11.724294

Figure Lengend Snippet: Ectopic GPC3 expression was successfully established and validated in isogenic SNU449 cells. ( A ) SNU449 cells were transduced with hGPC3-mGFP and clonally isolated, and GPC3 mRNA expression was quantified by qPCR. A431/GPC3 and HepG2 cells served as positive controls; A431 and GPC3⁻ HepG2 cells served as negative controls. ( B ) GPC3 protein expression was assessed by Western blotting (GAPDH loading control). ( C ) Cell-surface GPC3 was detected in A431/GPC3, SNU449/GPC3, and HepG2 cells, but not in GPC3⁻ SNU449, SNU449/vector, A431, or HepG2 cells. ( D ) Immunofluorescence staining confirmed GPC3 expression in SNU449/GPC3 and A431/GPC3 cells, but not in parental controls. Scale bars: 50 μ m.

Article Snippet: Human tissue specimens were incubated with a mouse monoclonal anti-human GPC3 antibody (clone 1G12; Cell Marque) under the same conditions.

Techniques: Expressing, Transduction, Isolation, Western Blot, Control, Plasmid Preparation, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Ablation of glypican-3 enhances radiosensitivity in liver cancer by prolonging G2/M arrest and activating the ATM/CHK2 pathway

doi: 10.64898/2026.05.11.724294

Figure Lengend Snippet: GPC3 expression regulates tumor cell proliferation and radiosensitivity. Proliferation of paired GPC3-positive (GPC3⁺) and GPC3-deficient (GPC3⁻) HepG2 ( A ), Hep3B ( B ), SNU449 and SNU449/V (empty vector control) ( C ), and A431 ( D ) cells was monitored using the IncuCyte® live-cell analysis system under non-irradiated (Non-IR) conditions and following 6 Gy irradiation (IR). Phase-area confluence was normalized to 0 h, and differences in proliferation kinetics were analyzed by two-way ANOVA, with significance assessed at the final time point (****, P <0.0001). Radiosensitivity was evaluated by clonogenic survival assays in the corresponding GPC3⁺ and GPC3⁻ HepG2 ( E ), Hep3B ( F ), SNU449 ( G ), and A431 ( H ) cells exposed to graded doses of γ-irradiation. Colonies were quantified 10–14 days later to generate survival curves. GPC3 loss increased radiosensitivity, yielding dose-modifying factors (DMFs) of 1.25 (HepG2), 1.36 (Hep3B), 1.20 (SNU449), and 1.75 (A431). Data represent mean surviving fraction ± SD from more than three independent experiments.

Article Snippet: Human tissue specimens were incubated with a mouse monoclonal anti-human GPC3 antibody (clone 1G12; Cell Marque) under the same conditions.

Techniques: Expressing, Plasmid Preparation, Control, Cell Analysis, Irradiation

Journal: bioRxiv

Article Title: Ablation of glypican-3 enhances radiosensitivity in liver cancer by prolonging G2/M arrest and activating the ATM/CHK2 pathway

doi: 10.64898/2026.05.11.724294

Figure Lengend Snippet: GPC3 deficiency increases IR-induced DNA double-strand breaks in tumor cells. DNA damage responses were assessed in paired GPC3-positive (GPC3⁺) and GPC3-deficient (GPC3⁻) HepG2 ( A , E ), Hep3B ( B , F ), SNU449 ( C , G ), and A431 ( D , I ) cells following 6 Gy irradiation (IR). γ-H2AX immunocytochemistry ( A – D ) was performed at the indicated time points to quantify DNA double-strand break formation and resolution. Representative images at control, 1 h, and 48 h post-IR are shown. γ-H2AX foci (magenta) were quantified from > 50 cells per condition; nuclei were counterstained with DAPI (blue). Scale bars: 25 μ m. Statistical comparisons of foci numbers were performed using a t -test (* p <0.05; *** p <0.001). Comet assays ( E – I ) were performed 24 h after IR. Representative comet images (left) and quantified tail moments (right) are shown. Data are presented as mean ± SD, and significance was determined using a t -test (** p <0.01; *** p <0.001; **** p <0.0001). Scale bars: 50 μ m.

Article Snippet: Human tissue specimens were incubated with a mouse monoclonal anti-human GPC3 antibody (clone 1G12; Cell Marque) under the same conditions.

Techniques: Irradiation, Immunocytochemistry, Control

Journal: bioRxiv

Article Title: Ablation of glypican-3 enhances radiosensitivity in liver cancer by prolonging G2/M arrest and activating the ATM/CHK2 pathway

doi: 10.64898/2026.05.11.724294

Figure Lengend Snippet: GPC3 deficiency enhances radiation response in xenograft models and correlates with clinical outcomes. ( A – D ) HepG2/Luc xenografts derived from GPC3⁺ or GPC3⁻ cells with or without 10 Gy irradiation were monitored by bioluminescence imaging (BLI) and tumor volume measurements. GPC3⁻ tumors showed enhanced growth delay following irradiation. Representative GPC3 and Ki-67 staining is shown (scale bar: 100 μ m). ( E – H ) Similar analyses in A431/Luc xenografts demonstrated greater radiation sensitivity in GPC3⁻ tumors. ( I ) In a clinical cohort, high tumor GPC3 expression was associated with poorer overall survival ( p = 0.008). Statistical comparisons were performed by two-way ANOVA; significance was assessed at the final time point (* p <0.05; *** p <0.001; **** p <0.0001).

Article Snippet: Human tissue specimens were incubated with a mouse monoclonal anti-human GPC3 antibody (clone 1G12; Cell Marque) under the same conditions.

Techniques: Derivative Assay, Irradiation, Imaging, Staining, Expressing

Journal: Frontiers in Immunology

Article Title: The CXCL9/SPP1 polarity axis in tumor-associated macrophages: immunoregulatory and prognostic significance in non-small cell lung cancer

doi: 10.3389/fimmu.2026.1763652

Figure Lengend Snippet: Validation of CXCL9 + and SPP1 + macrophage expression and polarization characteristics in lung cancer tissues. (A) Representative mIF staining for CXCL9+ TAMs and SPP1+ TAMs in lung tissue sections from the normal and LUAD tumor groups. DAPI (blue), CD68 (red), CXCL9 (yellow), SPP1 (green) are shown, along with individual and merged channels. (n = 3 per group). Scale bar, 20 μm. (B) Representative mIF staining of CXCL9+ TAMs (yellow) and SPP1+ TAMs (green) in lung tissue sections from the normal and LUAD tumor groups (n = 3 per group). Scale bar, 20 μm. (C) qRT-PCR validation of macrophage polarization. The M1 group showed high expression of Cxcl9 and iNOS, while the M2 group showed high expression of Spp1 and Arg1. Asterisks indicate statistically significant differences (*p < 0.05).

Article Snippet: Following another stripping step, the third round was performed using mouse anti-human CD68 monoclonal antibody (1:50000, Servicebio, Cat# GB153150 ) with HRP-conjugated goat anti-mouse secondary antibody (ready-to-use, Servicebio, Cat# G1301), and signal was developed using iF555-Tyramide (1:500, Servicebio, Cat# G1233).

Techniques: Biomarker Discovery, Expressing, Staining, Quantitative RT-PCR