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The plate was then incubated for ~30 min in the dark at room temperature with gentle orbital shaking

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IL-1 and TNF- released from microglia can open astrocytic hemichannels and reduce coupling between astrocytes in primary cell/slice cultures and in vivo in mice (Mme et al., 2006

Create a protocol checklist: baseline tests, informed consent, sourcing verification, monitoring schedule, stop rules for pregnancy, and documentation processes