Optogenetic Control of PIP2 Interactions Shaping ENaC Activity

Description

Comparison of the baseline Na+ levels by CoroNa Fluorescence and the membrane levels of ENaC before PIP2 depletion, under TIRF microscopy. Images were analyzed by image J and recorded the intensities.

Steps to reproduce

Create the PIP2 mutants on the sites described in the publication. Transfect HEK293T or CHO cells for 24-36 hours. HEK cells are used for imaging and CHO cells are used for patch clamp. For imaging, keep cells in the dark and then activate CIBN-CRY2 dimerization with Blue light briefly, then image under TIRF 10 min after. For patch clamp, start under light and then measure the activity under PIP2 recovery in the dark. More details are found in the publication.

Institutions

• The University of Texas Health Science Center at San Antonio

  Texas, San Antonio

Categories

Funders

• American Heart Association

  Dallas

  Grant ID: 857310
• American Heart Association

  Dallas

  Grant ID: 20POST35210746
• National Institute of Diabetes and Digestive and Kidney Diseases

  National Institutes of Health

  Bethesda

  Grant ID: DK113816

Licence