T22‐GFP‐H6‐FdU‐induced depletion of CXCR4‐overexpressing cancer cells in tumor tissue T22‐GFP‐H6‐FdU‐induced depletion of CXCR4‐overexpressing cancer cells.

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T22‐GFP‐H6‐FdU‐induced depletion of CXCR4‐overexpressing cancer cells in tumor tissue T22‐GFP‐H6‐FdU‐induced depletion of CXCR4‐overexpressing cancer cells in tumor tissue ARepresentative images of CXCR4 overexpression in subcutaneous tumor tissue, showing similar CXCR4 levels among compared groups (N = 5/group; Buffer, T22‐GFP‐H6‐FdU, T22‐GFP‐H6, and free oligo‐FdU) before treatment (upper panels). Representative images of DNA double‐strand break induction and caspase‐3 activation (measured with anti‐γ‐H2AX or anticleaved caspase‐3 by IHC) 5 h post‐administration (middle panels). Apoptotic induction (Hoechst staining, 24 h post‐administration, lower panels). Note the higher number of cells positive for DSBs, caspase‐3 activation, and apoptosis induction in the T22‐GFP‐H6‐FdU as compared to free oligo‐FdU. Black or white arrows indicate dead cells. Scale bar, 50 μm.BQuantitation of the number of cells containing DSBs or active caspase‐3 in IHC‐stained tumor sections 5 h post‐treatment and the number of condensated or disaggregated nuclei (by Hoechst staining) 24 h post‐treatment in tumor sections of 10 high‐power fields (400× magnification) using the Cell∧D software (N = 50; 10 tumor fields/mice; 5 mice/group). Data expressed as mean ± s.e.m. Parameter comparison between groups: (B: Buffer; T: T22‐GFP‐H6; F: free oligo‐FdU; T‐F: T22‐GFP‐H6‐FdU). P‐values for statistical differences: γ‐H2AX staining quantitation: B vs. T, #P = 0.001; B vs. F, #P = 0.000; B vs. T‐F, #P = 0.000; T vs. T‐F, **P = 0.001; F vs. T‐F, *P = 0.02. Cleaved caspase‐3 quantitation: B vs. F, *P = 0.034; B vs. T‐F, **P = 0.009; T vs. T‐F, **P = 0.003; F vs. T‐F, *P = 0.012. Hoechst staining quantitation: B vs. F, **P = 0.01; B vs. T‐F, **P = 0.001; T vs. T‐F, **P = 0.000; F vs. T‐F, *P = 0.032. Mann Whitney U‐test. María Virtudes Céspedes et al. EMBO Mol Med. 2018;emmm.201708772 © as stated in the article, figure or figure legend