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Fig. 1 | BMC Cancer

Fig. 1

From: Differential cytotoxicity induced by the Titanium(IV)Salan complex Tc52 in G2-phase independent of DNA damage

Fig. 1

Tc52 does not induce DNA damage. a: Detection of PAR (upper panel) and γH2AX (lower panel) by immunofluorescence after application of 10 μM Tc52 in HeLa cells. For positive controls (+), cells were incubated with 500 μM H2O2 for 10 min (PAR detection) and for 6 h (γH2AX detection). Respective upper rows show staining of nuclei (DAPI), lower rows detection of either PAR or γH2AX. Incubation time is depicted between both panels. γH2AX is only evident in apoptotic cells after 24 h. For each independent experiment, at least 100 cells were analyzed in technical duplicates. b Detection of PAR (upper panel) and γH2AX (lower panel) by immunofluorescence after application of 10 μM Tc52 in VH7 normal fibroblasts. For positive controls (+), cells were incubated with 500 μM H2O2 for 30 min (PAR detection) and for 1 h (γH2AX detection). Respective upper rows show staining of nuclei (DAPI), lower rows detection of either PAR or γH2AX. Incubation time is depicted between both panels. For each independent experiment, at least 100 cells were analyzed in technical duplicates. Complete time course of H2O2 treatment is depicted in Additional Fig. 3a for HeLa cells and Additional Fig. 3b for VH7 fibroblasts. c: Dose–response curve for mitomycin C-dependent crosslinking in HeLa. Bars indicate fluorescence signals with (filled bars) or without (open bars) 25 Gy irradiation as detected by FADU. Irradiation reduces, application of mitomycin C increases signals. Asterisks (*) indicate significant difference between irradiated and non-irradiated samples; hashtags (#) describe significance compared to the respective controls (0). p < 0.05 = #/*, p < 0.01 = ##/**; p < 0.001 = ###/***, (two-way ANOVA, Sidak's Multiple Comparison Test). At 100 μM mitomycin C, strong crosslinking prevents drop in signal intensity by irradiation. d: Time course of DNA breaks and crosslink detection by FADU after application of 10 μM Tc52 to HeLa cells for the indicated time points. Bars indicate fluorescence signals with (filled bars) or without (open bars) 25 Gy irradiation as detected by FADU. No change in signal intensity over time can be observed. 50 μg/ml mitomycinC (MMC) was applied as positive control. e: Time course of DNA breaks and crosslink detection by FADU after application of 10 μM Tc53 to HeLa cells for the indicated time points. Bars indicate fluorescence signals with (filled bars) or without (open bars) 25 Gy irradiation as detected by FADU. No change in signal intensity over time can be observed. 50 μg/ml mitomycinC (MMC) was applied as positive control

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