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Fig. 5 | Acta Neuropathologica Communications

Fig. 5

From: Astrocytes rescue neuronal health after cisplatin treatment through mitochondrial transfer

Fig. 5

Effects of cisplatin and astrocytes on neuronal calcium dynamics. Neurons were treated with cisplatin followed by co-culture with astrocytes and calcium dynamics were monitored with Fura-2. a, b. Resting ratio of 340:380 Fura-2 fluorescence intensity (F340:F380) as an indicator of intracellular levels of Ca2+ of neurons treated with or without cisplatin followed by co-culture with or without astrocytes. a. Mean traces and SEM of the F340:F380 ratio of Fura-2 at baseline during first 50 s of recording. b. Mean F340:F380 ratio during 50 s of recording for each neuron at rest. Data represents the mean ± SEM of > 90 neurons total per group collected in 3 independent experiments. One-way ANOVA, followed by Tukey’s post-hoc test: (*p < 0.05, ***p < 0.001, ****p < 0.0001). c. Mean traces and SEM of the increase in F340:F380 ratio of Fura-2 of neurons responding to stimulation with 20 mM KCl. The threshold change in Fura-2 F340:F380 ratio to be considered a response to 20 mM KCl was set at an increase of > 5 x standard deviations above baseline average. This excluded 9% of neurons in the control group, 23% of neurons in the cisplatin group and 11% of neurons in the cisplatin + astrocyte group. Data represents the mean ± SEM of 3 independent experiments with > 70 neurons for each group. d. Mean change in F340:F380ratio in response to stimulation of neurons with 20 mM KCl. The increase in Fura-2 ratio in response to 20 mM KCl was calculated for each neuron. Data represent the mean ± SEM of n > 90 neurons for each group collected in 3 independent experiments. One-way ANOVA, followed by Tukey’s post-hoc test: (**p < 0.01, ***p < 0.001, ****p < 0.0001). e Normalized KCl responses show a delayed decay of Ca2+ influx in cisplatin-treated neurons vs. control and cisplatin + astrocytes groups. f. Quantification of latency to return to 20% of maximal Ca2+ (dotted line). Data represents the mean ± SEM of 3 independent experiments with > 70 neurons for each group. One-way ANOVA, followed by Tukey’s post-hoc test: (*p < 0.05, ***p < 0.0001)

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