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In one aspect the disclosure sets forth a calcium oscillation assay as one way of measuring or predicting toxicity of a molecule. The disclosure also provides an in vivo tolerability assay that can be used separately or combined with the calcium oscillation assay and/or the sequence score method. FIG. 5: shows in vivo tolerability of exemplary antisense oligonucleotides. FIG. 4: shows the impact of Tau antisense oligonucleotide on spontaneous calcium oscillations in primary neurons. Effect of oligomers on primary neuronal spontaneous calcium oscillations was measured under two conditions in the presence and absence of 1 mM MgCl2 as described previously （Murphy et. Primary neuronal spontaneous calcium oscillations were measured as described previously （Murphy et. Networks of cortical neurons have been shown to undergo spontaneous calcium oscillations resulting in the release of the neurotransmitter glutamate. Regulated calcium oscillations are required for homeostasis of neuronal networks for normal brain |
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