Figure 6

Reducing neuronal activity mitigates the formation of DNA DSBs mediated by Dox.

(A) Dox upregulates Arc in neurons. Cortical neurons at 28–32 DIV were treated with a vehicle or with Dox (0.1 μM) overnight, fixed and stained for Arc (green) and imaged. Scale bar is 25 μm. (B) Images of fixed neurons from (A) were analysed. ***p < 0.0001 (t test). One hundred neurons were analysed. Results were pooled from three independent experiments. (C) Cortical neurons at 28–32 DIV were treated with a vehicle or with Dox (0.1 μM) or with Dox (0.1 μM) and TTX (1 μM), or with Dox (0.1 μM) and NBQX (50 μM), or with Dox (0.1 μM) and with APV (100 μM) overnight, fixed and stained for γH2A.X, MAP2c and with the nuclear Hoechst dye, imaged and analyzed with an automated algorithm. The puncta index was estimated by measuring the standard deviation of γH2A.X fluorescence intensity. *** p < 0.00001 (t-test). A.u., arbitrary units. Several thousand neurons were analyzed. Results were pooled from at least three independent experiments.