Modulation of Endocytic Pathways in Flow Cytometry

Immediately preceding insulin stimulation in flow cytometry preparations, cells were pre-treated with monodansylcadaverine (MDC) (20 μM, DMSO, Sigma), methyl-β-cyclodextrin (MβCD) (10 mM, culture media, Sigma), or relevant vehicle controls for 30 min at 37 °C [45 (link),46 (link),47 (link)]. MDC inhibits clathrin-mediated endocytosis by stabilizing clathrin-coated vesicles. MβCD disrupts caveolae by stripping cholesterol from the membrane and disrupting lipid rafts [48 (link)]. DMSO concentration was maintained at <0.5% in both experimental and vehicle treated samples.

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DiLucia S.G., Kendrick B.J, & Sims-Robinson C. (2023). Hyperinsulinemia Impairs Clathrin-Mediated Endocytosis of the Insulin Receptor and Activation of Endothelial Nitric Oxide Synthase in Brain Endothelial Cells. International Journal of Molecular Sciences, 24(19), 14670.

Publication 2023

DMSO culture media

Caveolae Cells Cholesterol Clathrin coated vesicles Culture media Cyclodextrin Dmso Endocytosis Flow cytometry Inhibits Insulin preparations Lipid Membrane Monodansylcadaverine

Corresponding Organization : Ralph H. Johnson VA Medical Center

Other organizations : MUSC Hollings Cancer Center

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Variable analysis

independent variables

Monodansylcadaverine (MDC) (20 μM, DMSO, Sigma)
Methyl-β-cyclodextrin (MβCD) (10 mM, culture media, Sigma)

dependent variables

Not explicitly mentioned

control variables

DMSO concentration was maintained at <0.5% in both experimental and vehicle treated samples
Relevant vehicle controls for 30 min at 37 °C

controls

Relevant vehicle controls for 30 min at 37 °C

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