Evaluation of Stable LifeAct-mRuby2- and LAMP1-NeonGreen Expressing A549 Cell Lines for Investigation of Interaction with Pulmonary Cells

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2021 Jun 2

PMID 34073107

Authors

Natalia Schiefermeier-Mach

Violetta Moresco

Stephan Geley

Lea Heinrich

Lukas Lechner

Heidi Oberhauser

Susanne Perkhofer

Affiliations

Soon will be listed here.

Abstract

Inhaled spores can be internalized by alveolar type II cells. Cell lines stably expressing fluorescently labeled components of endocytic pathway enable investigations of intracellular organization during conidia internalization and measurement of the process kinetics. The goal of this report was to evaluate the methodological appliance of cell lines for studying fungal conidia internalization. We have generated A549 cell lines stably expressing fluorescently labeled actin (LifeAct-mRuby2) and late endosomal protein (LAMP1-NeonGreen) following an evaluation of cell-pathogen interactions in live and fixed cells. Our data show that the LAMP1-NeonGreen cell line can be used to visualize conidia co-localization with LAMP1 in live and fixed cells. However, caution is necessary when using LifeAct-mRuby2-cell lines as it may affect the conidia internalization dynamics.

Citing Articles

Biological boundary conditions regulate the internalization of conidia by alveolar cells.

Schiefermeier-Mach N, Polleux J, Heinrich L, Lechner L, Vorona O, Perkhofer S Front Cell Infect Microbiol. 2025; 15:1515779.

PMID: 40066070 PMC: 11891256. DOI: 10.3389/fcimb.2025.1515779.

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