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. 2021 Jun 11;297(1):100873. doi: 10.1016/j.jbc.2021.100873

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© 2021 The Authors

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

PMC Copyright notice

Synj1 is expressed in the astrocytes and regulates astrocyte membrane P(4,5)P₂.A, the NBP1-87842 rabbit anti-Synj1 from Novus Biologicals recognizes the 145-kDa isoform of Synj1, which was abundantly expressed in the brain and weakly expressed in the astrocytes. White margin in the black box indicates the splicing border from the same membrane. B, Western blot analysis of Synj1 expression in adult mouse brain lysate, astrocyte lysate, microglia lysate, and HEK297T cell lysate as indicated using the NBP1-87842 polyclonal antibody. C, immunofluorescence for PI(4,5)P₂ in Synj1 WT, HET, and KO astrocytes sparsely transfected GFP-LC3. The PI(4,5)P₂ antibody was validated in our previous report (25). Membrane selections used for PI(4,5)P₂ analysis. D, analysis of the membrane PI(4,5)P₂ by tracing the contour of the transfected astrocytes shown in (C). p Value is from Tukey's post hoc test following one-way ANOVA. HEK297T, human embryonic kidney 297T; HET, heterozygous; Synj1, Synaptojanin1.