Fig. 5.

Phosphorylated VASP and phospho-null VASP protein–interacting networks.A, GFP-tagged human VASP (hVASP) WT and hVASP S239A are expressed in apoE4 Neuro-2a cells in triplicate in the absence or presence of 15 μM H89. GFP is used as a tag to pull down VASP protein–interacting partners which were identified using MS. B, heat map represents scaled protein abundance values for a selection of interacting proteins specifically regulated by hVASP S239 phosphorylation. C, comparison of hVASP WT with H89-treated hVASP WT protein interaction network for proteins displayed from panel B. (Red, increase; blue, decrease). D, bar plot of Gene Ontology analysis of proteins differentially purified from hVASP expressing apoE4 Neuro-2a cells in the absence or presence of H89. Proteins in Gene ontology analysis using EnrichR were determined by a p < 0.05 and a log₂ fold change >1 and sorted with p-value ranking. E, localization of VASP in apoE4 Neuro-2a cells in the absence or presence of 10 μM H89 for 24 h. Phalloidin staining is for F-actin. DAPI: nuclear staining. The scale bar is 10 μm. apo, apolipoprotein; hVASP, human VASP; MS, mass spectrometry; VASP, vasodilator-stimulated phosphoprotein.