Fig. 2

Evaluation of DNA damages induced by GO sheets after single exposure. A Quantification of lung DNA damages using rabbit anti-mouse γ-H2Ax-Phosphorilated Ser139 immunostaining after single exposure to 30 µg of GO sheets (USGO or LGO) and MWCNTs (positive control/reference materials) expressed as fold change in fluorescence intensity (negative control = water for injection). B Percentage of γ-H2Ax imunoreactivity (fluorescence intensity) located in lung immune cell infiltrates or in lung parenchyma at day 1, 7 and 28 after exposure. C Total number of γ-H2Ax positive cells in lung parenchyma (outside inflammatory infiltrates) expressed as fold change in fluorescence intensity. D Percentage of E-cadherin⁺ γ-H2Ax⁺ cells and CD45⁺ γ-H2Ax⁺ cells in lung parenchyma. E Representative images of lung section after DAPI staining and immunostaining (primary: rabbit anti-mouse γ-H2Ax-Phosphorilated Ser139; secondary: donkey anti-rabbit Alexa Fluor 647) for DNA damages. Mice were exposed by single oro-pharyngeal aspiration to a high dose GO sheets (USGO or LGO), MWCNTs, or water for injection. Scale bar = 50 µm. Significance level *p < 0.05 **p < 0.01, ***p < 0.001 (One-Way ANOVA; n = 3)