Recent Projects

Glycosylation Enzyme EXT1 Regulates ER Morphology and Cell Function

This study showed that the glycosyltransferase EXT1 plays a key role in shaping the structure and activity of the endoplasmic reticulum (ER) in mammalian cells. Using multi-omics and imaging, the team found that disrupting EXT1 affects ER shape, increases protein secretion, and alters cell metabolism, with implications for ER-linked organelles and cell development.

A Cytoskeletal Complex Links Actin, Microtubules, and ER Structure at Contact Sites

Insights into CDC20 and Chromosome Segregation in Plants

ER–Golgi Tubules Maintain Organelle Connectivity in Plant Cells

Engineering Plants to Help Reduce Methane Emissions

This study showed that the glycosyltransferase EXT1 plays a key role in shaping the structure and activity of the endoplasmic reticulum (ER) in mammalian cells. Using multi-omics and imaging, the team found that disrupting EXT1 affects ER shape, increases protein secretion, and alters cell metabolism, with implications for ER-linked organelles and cell development.

We identified a protein complex (NET3C–KLCR1–IQD2) that connects actin filaments and microtubules at ER–plasma membrane contact sites (EPCS) in plants. Loss of this complex disrupts cytoskeletal organization and ER morphology, revealing a critical link between cytoskeletal regulation and ER network maintenance.

This study characterized the plant homolog of CDC20, a key regulator of chromosome separation during mitosis. The team examined its structure, subcellular localization, and conservation across species, providing new insights into spindle checkpoint signaling in Arabidopsis and how it differs from other eukaryotes.

Methane is a powerful greenhouse gas, and rice farming alone contributes around 9% of global methane emissions. In this study, researchers took first steps toward creating rice plants that can break down methane before it escapes into the atmosphere. They focused on particulate methane monooxygenase (pMMO), a natural enzyme found in methane-eating bacteria.
The team successfully introduced and expressed the three pMMO subunits in plant models (tobacco and Arabidopsis), showing that these proteins could assemble correctly in plant cells. Although expression levels were low, this proof-of-concept shows that plants can potentially be engineered to detoxify methane, offering a novel strategy to reduce agricultural emissions.

Planning your next experiment

Working on exciting research? I’d love to collaborate and help with the latest microscopy techniques to give you clear, detailed insights for your next discovery. Here are some of the most recent techniques I have worked on.

Light sheet microscopy for imaging over long timescales

I used light sheet microscopy to capture high-resolution, long-term imaging of organelle dynamics in living plant cells, enabling observations over extended periods of up to 12 hours with minimal photodamage.

Electron microscopy

At Oxford Brookes University, I have access to high-pressure freezing and serial block-face scanning electron microscopy (SBF-SEM), enabling me to create highly detailed, three-dimensional images of plant organelles with exceptional structural preservation.

I used FRET-FLIM to characterise protein–protein interactions in living cells, allowing precise measurement of interaction dynamics in real time. This approach also enabled me to identify the topology of proteins within complexes, providing insight into their spatial arrangement and functional relationships.

FRET-FLIM

I used lipid dyes such as Rhodamine B to stain the endoplasmic reticulum, enabling detailed visualisation of ER structure in plant cells. This approach allowed clear imaging of the ER network and its organisation. By combining lipid staining with live-cell imaging, I could analyse ER morphology and its interactions with other organelles.

I used variable angle epifluorescence microscopy (VAEM) to track the movement of single particles within the plant secretory system. This technique enabled high-resolution imaging at the cell cortex, allowing detailed analysis of vesicle trafficking and dynamic interactions near the plasma membrane. Through VAEM, I was able to observe the behaviour of individual particles with exceptional spatial and temporal precision.

Lipid dyes

Variable angle epifluorescence microscopy (VAEM) for single particle tracking

Publication list

Studying ER-membrane contact sites in plants using the optogenetic approach: Taking the LiMETER as an example
Yifan Li, Charlotte Pain, Xuan Cui, Menghan Li, Tong Zhang, Jiejie Li, Verena Kriechbaumer, Pengwei Wang
The Plant Journal, 2025, 121(2): e17191

Overexpression of GFP Fusions of Regulators of the SAC from Arabidopsis thaliana
Charlotte Pain
Mitotic Exit, Springer US, 2024, 21-32

FRET-FLIM for the Study of Protein-Protein Interactions Underpinning Mitosis Checkpoints
Charlotte Pain
Mitotic Exit, Springer US, 2024, 87–97

Electron Microscopy Techniques for 3D Plant ER Imaging
Charlotte Pain and Maike Kittelmann
The Plant Endoplasmic Reticulum, Springer US, 2024, 15-25

Quantitation of ER Morphology and Dynamics
Mark Fricker, Emily Breeze Charlotte Pain, Verena Kriechbaumer, Carlos Aguilar, José M. Ugalde and Andreas J. Meyer
The Plant Endoplasmic Reticulum, Springer US, 2024, 49-75

Variable-Angle Epifluorescence Microscopy for Single-Particle Tracking in the Plant ER
Charlotte Pain, Christopher Tynan, Stanley W. Botchway, and Verena Kriechbaumer
The Plant Endoplasmic Reticulum, Springer US, 2024, 4273–2839-75

Observing ER Dynamics over Long Timescales Using Light Sheet Fluorescence Microscopy
Charlotte Pain, Verena Kriechbaumer and Alessia Candeo
The Plant Endoplasmic Reticulum, Springer US, 2024, 323–335

Recombinant expression and subcellular targeting of the particulate methane monooxygenase (pMMO) protein components in plants
Tatiana Spatola Rossi, Frances A. Tolmie, Tim Nichol, Charlotte Pain, Patrick Harrison, Thomas J. Smith, Mark Fricker and Verena Kriechbaumer
Scientific Reports, 2023, 13

In Depth Topological Analysis of Arabidopsis Mid-SUN Proteins and Their Interaction with the Membrane-Bound Transcription Factor MaMYB
Bisa Andov, Aurelia Boulaflous-Stevens, Charlotte Pain, Sarah Mermet, Maxime Voisin, Camille Charrondiere, Emmanuel Vanrobays, Sylvie Tutois, David E. Evans, Verena Kriechbaumer, Christophe Tatout and Katja Graumann
Plants, 2023, 9:1787

intER-ACTINg: The structure and dynamics of ER and actin are interlinked*
Charlotte Pain, Frances Tolmie, Stefan Wojcik, Pengwei Wang and Verena Kriechbaumer
Journal of Microscopy, 2022, 291:1(105-118)

*Top 10 most cited paper 2022
*Early Career Researcher Best Paper Award for 2023 in Life Sciences.

FRET-FLIM to Determine Protein Interactions and Membrane Topology of Enzyme Complexes
Tatiana Spatola Rossi, Charlotte Pain, Stanley W. Botchway and Verena Kriechbaumer
Current Protocols, 2022, 2:10

On the nature of the plant ER exit sites
Alastair J. McGinness, Jennifer Schoberer, Charlotte Pain, Federica Brandizzi and Verena Kriechbaumer
Frontiers in Plant Science, 2022, 13

Biochemical, biophysical, and functional characterisation of the E3 ubiquitin ligase APC/C regulator CDC20 from Arabidopsis thaliana
Maria-Alexa Cosma, Natalie L. Curtis, Charlotte Pain, Verena Kriechbaumer and Victor M. Bolanos-Garcia
Frontiers in Physiology, 2022, 13

Alternative glycosylation controls endoplasmic reticulum dynamics and tubular extension in mammalian cells
Despoina Kerselidou, Bushra Saeed Dohai, David R. Nelson, Sarah Daakour, Nicolas De Cock, et al… and Jean-Claude Twizere
Science Advances, 2021, 7:19

A novel plant actin-microtubule bridging complex regulates cytoskeletal and ER structure at ER-PM contact sites
Jingze Zang, Sandra Klemm, Charlotte Pain, Patrick Duckney, Zhiru Bao, Gina Stamm, Verena Kriechbaumer, Katharina BÃrstenbinder, Patrick J. Hussey and Pengwei Wang
Current Biology, 2021 31:6(1251-1260)

Living on the edge: the role of Atgolgin‐84A at the plant ER–Golgi interface
Vanessa Vieira, Charlotte Pain, Stephan Wojcik, Tatiana Spatola Rossi, Jurgen Denecke, Anne Osterrieder, Chris Hawes and Verena Kriechbaumer
Journal of Microscopy, 2020, 280(158-173)

Defining the dance: quantification and classification of endoplasmic reticulum dynamics
Charlotte Pain and Verena Kriechbaumer
Journal of Experimental Botany, 2019, 71:6(1757-1762)

Quantitative analysis of plant ER architecture and dynamics
Charlotte Pain, Verena Kriechbaumer, Maike Kittelmann, Chris Hawes and Mark Fricker
Nature Communications, 2019, 10

Arabidopsis Lunapark proteins are involved in ER cisternae formation
Verena Kriechbaumer, Emily Breeze, Charlotte Pain, Frances Tolmie, Lorenzo Frigerio and Chris Hawes
New Phytologist, 2018