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. 2022 Jul 16;3(3):101549. doi: 10.1016/j.xpro.2022.101549

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© 2022 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Immunofluorescence staining of PM PIPs and EEN1 in mouse hippocampal neurons

Mouse hippocampal neurons were transfected with pAOV-CaMKIIα-mCherry-2A-3Flag on DIV5 to express mCherry as volume marker (Note: the CaMKIIα promoter drives gene expression specifically in excitatory neurons but not inhibitory neurons) (Kohara et al., 2020), fixed on DIV8 and immnunostained for PM PI3P, PI4P or PI(4,5)P₂. DAPI-stained cell nuclei are shown in blue.

(A–D) Shown are representative confocal microscopy images of PM PI3P (A), PM PI4P (B), PM PI(4,5)P₂ (C), and costaining of PM PI(4,5)P₂ and PM EEN1 (D). Lower panels are magnifications of boxed regions in the top panels. Scale bars: 5 μm.