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Université Paris Sud

2 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Imaging Ca2+ Responses During Shigella Infection of Epithelial Cells
Yasmine Smail 1,2,3,4, Chunhui Sun 1,2,3,4, Laurent Combettes 5, Guy Tran Van Nhieu 1,2,3,4
1Equipe Communication Intercellulaire et Infections Microbiennes, Centre de Recherche Interdisciplinaire en Biologie (CIRB), Collège de France, 2Institut National de la Santé et de la Recherche Médicale (Inserm), U1050, 3Centre Nationale de la Recherche Scientifique (CNRS), UMR7241, 4MEMOLIFE Laboratory of Excellence and Paris Sciences et Lettres, 5Inserm, UMRS1174, Université Paris Sud

Here, we present protocols to visualize calcium (Ca2+) responses elicited by HeLa cells infected by Shigella. By optimizing the parameters of bacterial infection and imaging with Ca2+ fluorescent probes, atypical global and local Ca2+ signals induced by bacteria over a large range of infection kinetics are characterized.

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Biochemistry

Purification of Tubulin with Controlled Posttranslational Modifications and Isotypes from Limited Sources by Polymerization-Depolymerization Cycles
Satish Bodakuntla *1,2, A.S. Jijumon *1,2, Carsten Janke 1,2, Maria M. Magiera 1,2
1PSL Research University, CNRS UMR3348, Institut Curie, 2Université Paris-Saclay, CNRS UMR3348, Université Paris Sud

This protocol describes tubulin purification from small/medium-scale sources such as cultured cells or single mouse brains, using polymerization and depolymerization cycles. The purified tubulin is enriched in specific isotypes or has specific posttranslational modifications and can be used in in vitro reconstitution assays to study microtubule dynamics and interactions.

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