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Action of cetuximab-vc-MMAF on cell viability. A. Flow cytometry-based apoptosis assay of cells treated with the cetuximab-vc-MMAF (10 nM) for the indicated times. Live cells are shown in the lower left quadrant (Annexin V-/PI-), while the other three quadrants represent different stages of non-viable cells. B. Graphical representation of non-viable cells for each condition. Data are plotted as mean ± SD of three independent experiments. C. Western blot analysis of apoptosis-related proteins at the indicated time points following treatment with 10 nM cetuximab-vc-MMAF. Actin was used as a loading control, and molecular weight markers are shown at the right. D. Analysis of caspase-3 activity in cells treated with cetuximab-vc-MMAF (10 nM) for the indicated times. Data represent mean ± SD of fluorescence intensity resulting from substrate cleavage <t>(Ac-DEVD-AFC)</t> by activated caspase-3 in two independent experiments.
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Action of cetuximab-vc-MMAF on cell viability. A. Flow cytometry-based apoptosis assay of cells treated with the cetuximab-vc-MMAF (10 nM) for the indicated times. Live cells are shown in the lower left quadrant (Annexin V-/PI-), while the other three quadrants represent different stages of non-viable cells. B. Graphical representation of non-viable cells for each condition. Data are plotted as mean ± SD of three independent experiments. C. Western blot analysis of apoptosis-related proteins at the indicated time points following treatment with 10 nM cetuximab-vc-MMAF. Actin was used as a loading control, and molecular weight markers are shown at the right. D. Analysis of caspase-3 activity in cells treated with cetuximab-vc-MMAF (10 nM) for the indicated times. Data represent mean ± SD of fluorescence intensity resulting from substrate cleavage (Ac-DEVD-AFC) by activated caspase-3 in two independent experiments.

Journal: Neoplasia (New York, N.Y.)

Article Title: Rational payload selection enables high antitumoral efficacy of an anti-EGFR antibody-drug conjugate against ovarian tumors

doi: 10.1016/j.neo.2026.101295

Figure Lengend Snippet: Action of cetuximab-vc-MMAF on cell viability. A. Flow cytometry-based apoptosis assay of cells treated with the cetuximab-vc-MMAF (10 nM) for the indicated times. Live cells are shown in the lower left quadrant (Annexin V-/PI-), while the other three quadrants represent different stages of non-viable cells. B. Graphical representation of non-viable cells for each condition. Data are plotted as mean ± SD of three independent experiments. C. Western blot analysis of apoptosis-related proteins at the indicated time points following treatment with 10 nM cetuximab-vc-MMAF. Actin was used as a loading control, and molecular weight markers are shown at the right. D. Analysis of caspase-3 activity in cells treated with cetuximab-vc-MMAF (10 nM) for the indicated times. Data represent mean ± SD of fluorescence intensity resulting from substrate cleavage (Ac-DEVD-AFC) by activated caspase-3 in two independent experiments.

Article Snippet: The payloads used for antibodies conjugation, MC-GGFG-DXD (DXd, catalog reference: HY-13631E), MC-Val-Cit-PAB-MMAF (Vc-MMAF, catalog reference: HY-112786) and McMMAF (catalog reference: HY-15578), the belantamab mafodotin ADC (catalog reference: HY-P3239), as well as the Ac-DEVD-AFC (catalog reference: HY-P1005) substrate for caspase-3 activity detection, were purchased from MedChem Express (Princeton, NJ, USA).

Techniques: Flow Cytometry, Apoptosis Assay, Western Blot, Control, Molecular Weight, Activity Assay, Fluorescence