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Published in final edited form as: FEBS J. 2015 Nov 6;283(1):112–129. doi: 10.1111/febs.13556

Inhibition of DSB repair by anti-A3G peptides. Cells were irradiated (4 Gy) or mock-irradiated (No-IR) and stained with anti-A3G and anti-γ-H2AX antibody. Nuclei stained by DAPI (Blue); A3G stained by Alexa fluor 647 (red); γH2AX foci by Alexa fluor 488 (Green). (A) H9 and SupT1 cells before and 24h post-irradiation. (B) Non-irradiated H9 and SupT1 cells were cultivates in the presence of indicated peptides. (C) H9 and SupT1 cells were pre-incubated for 2 hours with the indicated peptides, irradiated (4 Gy) and stained after 24 hours. Top panels (from left): H9 cells in the presence of peptides: Mutant of Vif107-115_(110AA111); Vif30-39 (Negative control) and A3F304-312. Middle panels (from left): H9 cells irradiated in the presence of peptides, Vif25-39; Vif107-115 and A3F305-311. Bottom panels (from left): SupT1 cells in the presence of peptides: Mutant of Vif107-115_(110AA111); Vif25-39 and Vif107-115. (D) Ly-4 cells were pre-incubated for 2 hours with the indicated peptides, irradiated (4 Gy) or mock-irradiated (NoIR), and stained after 24 hours. Top panels (from left): Ly-4 cells non-irradiated; Ly-4 cells irradiated in the presence of peptides, Vif25-39; Vif107-115. Bottom panels (from left): Ly-4 cells irradiated; Ly-4 cells irradiated in the presence of peptides, A3F305-311 and A3F304-312. (E) Quantification of γ-H2AX foci in H9, Ly-4 and SupT1 24 hours post-IR (4 Gy). Values represent mean SD from two independent experiments in each at least five different microscopic fields were analyzed. * P < 0.05, ** P < 0.01, ANOVA, n = 50.

Inhibition of DSB repair by anti-A3G peptides. Cells were irradiated (4 Gy) or mock-irradiated (No-IR) and stained with anti-A3G and anti-γ-H2AX antibody. Nuclei stained by DAPI (Blue); A3G stained by Alexa fluor 647 (red); γH2AX foci by Alexa fluor 488 (Green). (A) H9 and SupT1 cells before and 24h post-irradiation. (B) Non-irradiated H9 and SupT1 cells were cultivates in the presence of indicated peptides. (C) H9 and SupT1 cells were pre-incubated for 2 hours with the indicated peptides, irradiated (4 Gy) and stained after 24 hours. Top panels (from left): H9 cells in the presence of peptides: Mutant of Vif107-115_(110AA111); Vif30-39 (Negative control) and A3F304-312. Middle panels (from left): H9 cells irradiated in the presence of peptides, Vif25-39; Vif107-115 and A3F305-311. Bottom panels (from left): SupT1 cells in the presence of peptides: Mutant of Vif107-115_(110AA111); Vif25-39 and Vif107-115. (D) Ly-4 cells were pre-incubated for 2 hours with the indicated peptides, irradiated (4 Gy) or mock-irradiated (NoIR), and stained after 24 hours. Top panels (from left): Ly-4 cells non-irradiated; Ly-4 cells irradiated in the presence of peptides, Vif25-39; Vif107-115. Bottom panels (from left): Ly-4 cells irradiated; Ly-4 cells irradiated in the presence of peptides, A3F305-311 and A3F304-312. (E) Quantification of γ-H2AX foci in H9, Ly-4 and SupT1 24 hours post-IR (4 Gy). Values represent mean SD from two independent experiments in each at least five different microscopic fields were analyzed. * P < 0.05, ** P < 0.01, ANOVA, n = 50.