Fig. 1

Astrocytes improve survival of neurons damaged by cisplatin. a-d. Survival of neurons (a, b) and astrocytes (c, d) after exposure to cisplatin. Primary cultures of cortical neurons (12 days in vitro) or cortical astrocytes were treated with increasing doses of cisplatin or vehicle for 24 h, then the cisplatin was removed and replaced with media. Survival was measured using a WST-1 assay: (a, c) at the end of the 24 h culture period or (b, d) 17 h after removal of cisplatin. Data was normalized to survival in the absence of cisplatin and analyzed using one-way ANOVA followed by Dunnett’s multiple comparisons test (*p < 0.05, **p < 0.01, ***p < 0.001). e. Astrocytes were treated with 1 μM cisplatin for 24 h followed by 17 h of culture without cisplatin. Oxygen consumption rates (OCR) were analyzed using Seahorse XFe 24 Analyzer and normalized to protein content. f. Neurons and astrocytes were treated with cisplatin or control medium for 24 h. Cisplatin was removed, neurons were labeled with Celltracker Blue (CTB) and cultured for another 17 h with or without astrocytes, and the number of surviving neurons was counted. Data are normalized to control and represents the mean ± SEM of 3 independent experiments performed in duplicate. Two-way ANOVA: cisplatin x astrocyte interaction: **p < 0.01; Tukey’s multiple comparisons post hoc test (*p < 0.05, **p < 0.01, ****p < 0.0001)