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. 2021 Oct 27;15(1):101257. doi: 10.1016/j.tranon.2021.101257

Fig. 1.

Fig 1

Target identification, localization, and specificity of mAb150 in A4 cells. A-i. Immunoblotting of A4 whole cell lysates with mAb150; A-ii,A-iii. Cross-immunoprecipitation (IP) followed by probing of A4 lysate and Ax1 protein with mAb150 and commercial AnxA2 antibody and relevant controls; A-iv, A-v. Representative IB of mAb150 and commercial AnxA2 antibody with pure human recombinant Anxa2 protein and A4 lysates; A-vi, Avii. Localization of mAb150 target across membrane (Mem), cytoplasm (Cyto) and nuclear (Nuc) subcellular fractions (CD24, B-actin and H3A are respective controls) in A4 cells; B-i, ii. Anxa2 expression profiling through immunofluorescence staining and flow cytometry respectively; B-iii, B-iv. Cytotoxicity of mAb150 in A4 cells (MTT) and xenografts respectively; B-v. Significant reduction of AnxA2 expressing tumor cells following six doses of mAb150 treatment (18 mg/Kg) with 4 days of recovery between consecutive doses. For all experiments and wherever relevant, IgM isotype control (Sigma-Aldrich # M-5909) was used. *P<0.05; **P<0.01; ***P<0.001.