Institut Pasteur Shanghai and Chinese Academy of Sciences

2 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Immunofluorescence Analysis of Stress Granule Formation After Bacterial Challenge of Mammalian Cells

Pascale Vonaesch¹, Philippe J. Sansonetti^1,2, Pamela Schnupf³

¹Unité de Pathogénie Microbienne Moléculaire, INSERM U786, Institut Pasteur, ²Microbiologie et Maladies Infectieuses, Collège de France, ³Laboratory of Intestinal Immunity, Institut Imagine-INSERM UMR 1163, Université Paris Descartes-Sorbonne Paris Cité

We describe a method for the qualitative and quantitative analysis of stress granule formation in mammalian cells after the cells are challenged with bacteria and a number of different stresses. This protocol can be applied to investigate the cellular stress granule response in a wide range of host-bacterial interactions.

Biology

Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe

Aline Stedman^1,3, Antonin Levy^1,4, Philippe J. Sansonetti^1,2,5, Giulia Nigro^1,6

¹Molecular Microbial Pathogenesis Unit, Institut Pasteur, ²Chaire de Microbiologie et Maladies Infectieuses, Collège de France, ³Institut de Biologie Paris Seine (IBPS) - Developmental Biology Unit, Sorbonne Université, CNRS UMR7622, INSERM U1156, ⁴Molecular Radiotherapy, INSERM U1030, Gustave Roussy, Université Paris-Saclay, ⁵The Center for Microbes, Development and Health, Institut Pasteur Shanghai and Chinese Academy of Sciences, ⁶Microenvironment and Immunity Unit, Institut Pasteur

The present protocol describes a method to detect reactive oxygen species (ROS) in the intestinal murine organoids using qualitative imaging and quantitative cytometry assays. This work can be potentially extended to other fluorescent probes to test the effect of selected compounds on ROS.