Marc Tramier
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Protocol for quantifying LC3B FRET biosensor activity in living cells using a broad-to-sensitive data analysis pipeline Open
Here, we present a protocol to comprehensively quantify autophagy initiation using the readout of the microtubule associated protein 1 light chain 3 beta (LC3B) Förster's resonance energy transfer (FRET) biosensor. We describe steps for ce…
View article: Actin cytoskeleton differently regulates cell surface organization of GPI-anchored proteins in polarized epithelial cells and fibroblasts
Actin cytoskeleton differently regulates cell surface organization of GPI-anchored proteins in polarized epithelial cells and fibroblasts Open
The spatiotemporal compartmentalization of membrane-associated glycosylphosphatidylinositol-anchored proteins (GPI-APs) on the cell surface regulates their biological activities. These GPI-APs occupy distinct cellular functions such as enz…
The LC3B FRET biosensor monitors the modes of action of ATG4B during autophagy in living cells Open
Although several mechanisms of macroautophagy/autophagy have been dissected in the last decade, following this pathway in real time remains challenging. Among the early events leading to its activation, the ATG4B protease primes the key au…
The LC3B FRET biosensor monitors the modes of action of ATG4B during autophagy in living cells Open
Although several mechanisms of macroautophagy/autophagy have been dissected in the last decade, following this pathway in real time remains challenging. Among the early events leading to its activation, the ATG4B protease primes the key au…
The LC3B FRET biosensor monitors the modes of action of ATG4B during autophagy in living cells Open
Although several mechanisms of autophagy have been dissected in the last decade, following this pathway in real time remains challenging. Among the early events leading to its activation, the ATG4B protease primes the key autophagy player …
Neural network fast‐classifies biological images through features selecting to power automated microscopy Open
Artificial intelligence is nowadays used for cell detection and classification in optical microscopy during post‐acquisition analysis. The microscopes are now fully automated and next expected to be smart by making acquisition decisions ba…
Label-free microscopy of mitotic chromosomes using the polarization orthogonality breaking technique Open
We report how a recently developed polarization imaging technique, implementing micro-wave photonics and referred to as orthogonality-breaking (OB) imaging, can be adapted on a classical confocal fluorescence microscope, and is able to pro…
View article: tdLanYFP, a yellow, bright, photostable and pH insensitive fluorescent protein for live cell imaging and FRET-based sensing strategies
tdLanYFP, a yellow, bright, photostable and pH insensitive fluorescent protein for live cell imaging and FRET-based sensing strategies Open
Yellow fluorescent proteins (YFP) are widely used as optical reporters in Förster Resonance Energy Transfer (FRET) based biosensors. Although great improvements have been done, the sensitivity of the biosensors is still limited by the low …
Mitochondrial Aurora kinase A induces mitophagy by interacting with MAP1LC3 and Prohibitin 2 Open
Epithelial and haematologic tumours often show the overexpression of the serine/threonine kinase AURKA. Recently, AURKA was shown to localise at mitochondria, where it regulates mitochondrial dynamics and ATP production. Here we define the…
View article: The NANOTUMOR consortium – Towards the Tumor Cell Atlas
The NANOTUMOR consortium – Towards the Tumor Cell Atlas Open
Cancer is a multi‐step disease where an initial tumour progresses through critical steps shaping, in most cases, life‐threatening secondary foci called metastases. The oncogenic cascade involves genetic, epigenetic, signalling pathways, in…
Neural network fast-classifies biological images using features selected after their random-forests-importance to power smart microscopy Open
Artificial intelligence is nowadays used for cell detection and classification in optical microscopy, during post-acquisition analysis. The microscopes are now fully automated and next expected to be smart, to make acquisition decisions ba…
Neural network fast-classifies biological images using features selected after their random-forests-importance to power smart microscopy Open
Artificial intelligence is nowadays used for cell detection and classification in optical microscopy, during post-acquisition analysis. The microscopes are now fully automated and next expected to be smart, to make acquisition decisions ba…
Label-free microscopy of mitotic chromosomes using the polarization Orthogonality Breaking technique Open
The vast majority of the microscopy methods currently available to study biological samples require staining prior to imaging. Nevertheless, the ability to reveal specific cell structures or organelles in a label-free manner remains desira…
Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells Open
Epithelial cancers are often hallmarked by the overexpression of the Ser/Thr kinase Aurora A/AURKA. AURKA is a multifunctional protein that activates upon its autophosphorylation on Thr288. AURKA abundance peaks in mitosis, where it contro…
Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells Open
Epithelial cancers are often hallmarked by the overexpression of the Ser/Thr kinase Aurora A/AURKA. AURKA is a multifunctional protein that activates upon its autophosphorylation on Thr288. AURKA abundance peaks in mitosis, where it contro…
Mitochondrial Aurora kinase A induces mitophagy by interacting with MAP1LC3 and Prohibitin 2 Open
Epithelial and haematologic tumours often show the overexpression of the serine/threonine kinase AURKA. Recently, AURKA was shown to localise at mitochondria, where it regulates mitochondrial dynamics and ATP production. Here we define the…
Automated screening of AURKA activity based on a genetically encoded FRET biosensor using fluorescence lifetime imaging microscopy Open
Fluorescence Lifetime Imaging Microscopy (FLIM) is a robust tool to measure Förster Resonance Energy Transfer (FRET) between two fluorescent proteins, mainly when using genetically-encoded FRET biosensors. It is then possible to monitor bi…
Imaging Mitochondrial Functions: From Fluorescent Dyes to Genetically-Encoded Sensors Open
Mitochondria are multifunctional organelles that are crucial to cell homeostasis. They constitute the major site of energy production for the cell, they are key players in signalling pathways using secondary messengers such as calcium, and…
View article: Optimized FRET Pairs and Quantification Approaches To Detect the Activation of Aurora Kinase A at Mitosis
Optimized FRET Pairs and Quantification Approaches To Detect the Activation of Aurora Kinase A at Mitosis Open
Genetically encoded Förster's Resonance Energy Transfer (FRET) biosensors are indispensable tools to sense the spatiotemporal dynamics of signal transduction pathways. Investigating the crosstalk between different signaling pathways is bec…
Chromatin condensation fluctuations rather than steady-state predict chromatin accessibility Open
Chromatin accessibility to protein factors is critical for genome activities. However, the dynamic properties of chromatin higher-order structures that regulate its accessibility are poorly understood. Here, we took advantage of the microe…
View article: Optimised FRET pairs and quantification approaches to detect the activation of Aurora kinase A at mitosis
Optimised FRET pairs and quantification approaches to detect the activation of Aurora kinase A at mitosis Open
Genetically-encoded Förster’s Resonance Energy Transfer (FRET) biosensors are indispensable tools to sense the spatiotemporal dynamics of signal transduction pathways. Investigating the crosstalk between different signalling pathways is be…
The polarity-induced force imbalance in<i>Caenorhabditis elegans</i>embryos is caused by asymmetric binding rates of dynein to the cortex Open
During asymmetric cell division, the molecular motor dynein generates cortical pulling forces that position the spindle to reflect polarity and adequately distribute cell fate determinants. In Caenorhabditis elegans embryos, despite a meas…
Aurora kinase A localises to mitochondria to control organelle dynamics and energy production Open
Many epithelial cancers show cell cycle dysfunction tightly correlated with the overexpression of the serine/threonine kinase Aurora A (AURKA). Its role in mitotic progression has been extensively characterised, and evidence for new AURKA …
Author response: Aurora kinase A localises to mitochondria to control organelle dynamics and energy production Open
Article Figures and data Abstract eLife digest Introduction Results Discussion Materials and methods Data availability References Decision letter Author response Article and author information Metrics Abstract Many epithelial cancers show …
Chromatin Condensation Fluctuations Rather than Steady-State Predict Chromatin Accessibility Open
Chromatin accessibility to protein factors is critical for genome activities. Dynamic changes in nucleosomal DNA compaction and higher order chromatin structures are expected to allow specific sites to be accessible to regulatory factors a…