Monitoring of Autophagy and Cell Volume Regulation in Kidney Epithelial Cells in Response to Fluid Shear Stress

Methods Mol Biol. 2019:1880:331-340. doi: 10.1007/978-1-4939-8873-0_22.

Abstract

Fluidic shear stress applied to epithelial cells inside the kidney tubules affects cell size in an autophagy-related manner. Here, we describe the technical equipment that we routinely use to apply shear stress on cells, as well as immunoblotting, immunofluorescence, and three-dimensional cell volume reconstruction techniques used in analysis of the influence of this stress on cells and cellular components. By pointing out details of experimental techniques and potential pitfalls, this review will serve as a guide for those interested in study of how shear stress influences cells.

Keywords: Autophagy; Cell volume; Fluid flow; Immunoblot; Immunofluorescence; Microscopy; Microslides; Primary cilia; Shear stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autophagy / physiology*
  • Biological Assay / instrumentation
  • Biological Assay / methods*
  • Cell Line
  • Cell Size*
  • Dogs
  • Epithelial Cells / cytology*
  • Epithelial Cells / physiology
  • Humans
  • Imaging, Three-Dimensional / instrumentation
  • Imaging, Three-Dimensional / methods*
  • Mice
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Shear Strength / physiology
  • Software
  • Stress, Mechanical