Ann Wheeler

• PhD Cell Biology, UCL
• Head of Department at University of Edinburgh

Inappropriate stimulation or defective negative regulation of the type I interferon response can lead to autoinflammation. In genetically uncharacterized cases of the type I interferonopathy Aicardi–Goutières syndrome, we identified biallelic mutations in LSM11 and RNU7-1, which encode components of the replication-dependent histone pre-mRNA–proces...

In the version of this paper originally published, Figure 4a contained errors that were introduced during typesetting. The bottom 11° ThunderSTORM image is an xz view but was incorrectly labeled as xy, and the low x-axis value in the four line profiles was incorrectly set as –60 instead of –50. These errors have been corrected in the PDF and HTML v...

With the widespread uptake of two-dimensional (2D) and three-dimensional (3D) single-molecule localization microscopy (SMLM), a large set of different data analysis packages have been developed to generate super-resolution images. In a large community effort, we designed a competition to extensively characterize and rank the performance of 2D and 3...

Source Data for Expanded View

Expanded View Figures PDF

During apoptosis, pro-apoptotic BAX and BAK are activated, causing mitochondrial outer membrane permeabilisation (MOMP), caspase activation and cell death. However, even in the absence of caspase activity, cells usually die following MOMP Such caspase-independent cell death is accompanied by inflammation that requires mitochondrial DNA (mtDNA) acti...

With the widespread uptake of 2D and 3D single molecule localization microscopy, a large set of different data analysis packages have been developed to generate super-resolution images. To guide researchers on the optimal analytical software for their experiments, we have designed, in a large community effort, a competition to extensively character...

Super-resolution microscopy methods enable resolution of biological molecules in their cellular or tissue context at the nanoscale. Different methods have their strengths and weaknesses. Here we present a method that enables correlative confocal, structured illumination microscopy (SIM) and single-molecule localization microscopy (SMLM) imaging of...

During apoptosis, pro-apoptotic BAX and BAK are activated, causing mitochondrial outer membrane permeabilisation (MOMP), caspase activation and cell death. However, even in the absence of caspase activity, cells usually die following MOMP. Such caspase-independent cell death is accompanied by inflammation that requires mitochondrial DNA (mtDNA) act...

Table S1: Summary of validated de novo variants identified in UK10K Trio analyses

Table S2: Summary of quantitative heavy vs light labelled mass spectrometry data from GFP‐TRAP Co‐immunoprecipitates

Supporting Information

DNA is strictly compartmentalized within the nucleus to prevent autoimmunity; despite this, cyclic GMP-AMP synthase (cGAS), a cytosolic sensor of double-stranded DNA, is activated in autoinflammatory disorders and by DNA damage. Precisely how cellular DNA gains access to the cytoplasm remains to be determined. Here, we report that cGAS localizes to...

Ocular coloboma (OC) is a defect in optic fissure closure and is a common cause of severe congenital visual impairment. Bilateral OC is primarily genetically determined and shows marked locus heterogeneity. Whole exome sequencing was used to analyse twelve trios (child affected with OC and both unaffected parents), This identified de novo mutations...

There was an error published in J. Cell Sci. 125 , [3153–3163][1]. In [Fig. 7][2]A, the MDA-MB-231 shPLCγ1+GSK2334470 sample was mislabelled as shPLCγ1+GSK2337740. Additionally, that image was inadvertently duplicated as the A375M shPLCγ1 image in panel B. The correct [Fig. 7][2] is shown

We demonstrate a spectroscopic imaging based super-resolution approach by separating the overlapping diffraction spots into several detectors during a single scanning period and taking advantage of the size-dependent emission wavelength in nanoparticles. This approach has been tested using off-the-shelf quantum dots (Invitrogen Qdot) and in-house n...

Nature Communications 7 : Article number: 13542 10.1038/ncomms13542 ( 2016 ); Published: 6 December 2016 ; Updated: 5 January 2017 The original version of this Article contained an error in the spelling of the author Tommaso Poggioli, which was incorrectly given as Tommaso Pogglioli.

In spite of extensive recent progress, a comprehensive understanding of how actin cytoskeleton remodelling supports stable junctions remains to be established. Here we design a platform that integrates actin functions with optimized phenotypic clustering and identify new cytoskeletal proteins, their functional hierarchy and pathways that modulate E...

Supplementary Figures 1-9, Supplementary Tables 1-3, Supplementary Methods and Supplementary References.

Gillespie syndrome (GS) is characterized by bilateral iris hypoplasia, congenital hypotonia, non-progressive ataxia, and progressive cerebellar atrophy. Trio-based exome sequencing identified de novo mutations in ITPR1 in three unrelated individuals with GS recruited to the Deciphering Developmental Disorders study. Whole-exome or targeted sequence...

[This corrects the article DOI: 10.1371/journal.pgen.1005673.].

Human β-defensin 3 (hBD3) is a cationic host defence peptide and is part of the innate immune response. HBD3 is present on a highly copy number variable block of six β-defensin genes, and increased copy number is associated with the autoimmune disease psoriasis. It is not known how this increase influences disease development, but psoriasis is a T...

Free-standing Ge quantum dots around 3 nm in size were synthesized using a bench-top colloidal method and suspended in water and ethanol. In the ethanol solution, the photoluminescence of the Ge quantum dots was observed between 650 and 800 nm. Structural and optical properties of these colloidal Ge quantum dots were investigated by utilizing X-ray...

Toll-like receptor 9 (TLR9) has a key role in the recognition of pathogen DNA in the context of infection and cellular DNA that is released from damaged cells. Pro-inflammatory TLR9 signalling pathways in immune cells have been well investigated, but we have recently discovered an alternative pathway in which TLR9 temporarily reduces energy substra...

In this study we use a spinning disk confocal microscope (SD) to generate super-resolution images of multiple cellular features from any plane in the cell. We obtain super-resolution images by using stochastic intensity fluctuations of biological probes, combining Photoactivation Light-Microscopy (PALM)/Stochastic Optical Reconstruction Microscopy...

The potential to use Schwann cells (SCs) in neural repair for patients suffering from neurotrauma and neurodegenerative diseases is well recognized. However, significant cell death after transplantation hinders the clinical translation of SC-based therapies. Various factors may contribute to the death of transplanted cells. It is known that prolong...

The question of whether the distinct isoforms of the family of enzymes phosphoinositide 3-kinases (PI3Ks) play redundant roles within a cell or whether they control distinct cellular processes or distinct steps within the same cellular process has gained considerable importance in the recent years due to the development of inhibitors able to select...

Effect of PI3K inhibitors on S1P- and HDL3-induced EC migration. Serum-starved HUVEC were pre-treated with 1 µM LY294002 (A), 5 µM LY294002 (B), 10 µM LY294002 (C), 1 µM AS605240 (D) or 100 nM of the specific p110β inhibitor TGX221 (D) for 30 min. Cell migration induced by S1P was determined by Transwell assays. Data are expressed as percentage of...

Class II and class IB PI3Ks are involved in remodelling of HUVEC. (A-C) Results from analysis of EC rearrangement. Data indicate the total number of branching points and are means ± SEM from 3–4 (A), 3 (B), and 4 (C) independent experiments. (D,E) The S1P- and HDL3-dependent HUVEC was assessed in the absence or presence of 1 µM A66. Representative...

Primers used for RT-PCR analysis presented in Figure S3. (DOC)

Effect of PI3Ks downregulation and PI3K inhibitors on the S1P-dependent Akt and ERK phosphorylation. Representative images of Western blotting analysis of Akt and ERK phosphorylation performed in HUVEC. Membranes were then stripped and incubated with the corresponding total antibodies. (A-B) HUVEC transfected with the indicated siRNAs were serum st...

Class II and class IB PI3Ks are involved in S1P-induced EC migration. (A) Results from Transwell assays performed in control, untransfected HUVEC and HUVEC transfected with a scrambled siRNA. Data are expressed as percentage of control (cells untransfected and unstimulated) and are means ± SEM from 6 independent experiments. (B) Levels of PI3K-C2α...

RT-qPCR analysis of PI3Ks levels. (A) Downregulation of p110γ mRNA levels using two distinct siRNAs was determined by RT-qPCR. (B) HUVEC were transfected with siRNAs targeting the indicated PI3Ks. Efficiency and specificity of downregulation was determined by RT-qPCR. (TIF)

Effect of PI3Ks downregulation on EC apoptosis. (A) Results from caspase 3 assay performed on lysates from HUVEC obtained 48 h after transfection with the indicated siRNAs (sequences 2). Data are means ± SEM from 2 independent experiments. (B) HUVEC transfected with the indicated siRNAs were incubated in serum free M119 after 24 h from transfection...

S1P induces de novo synthesis of PtdIns3P. HUVEC were labelled with [3H]myo-inositol before stimulation with 1 µM S1P for the indicated times. Phosphoinositides were then extracted, deacylated and analysed by HPLC. Data show levels of glyceroPtdIns3P normalised for the levels of glycerophosphatidylinositol (gPtdIns) and expressed as percentage of P...

The epidermis and its appendages, the hair follicle and sebaceous gland, have the capacity to constantly regenerate throughout adult life. Postnatal hair follicles undergo a cyclic mode of tissue homeostasis, defined by periods of growth, degeneration, and rest. A multipotent population of stem cells residing within the hair follicle bulge not only...

Desmoglein 3 (Dsg3), a member of the desmoglein sub-family, serves as an adhesion molecule in desmosomes. Our previous study showed that overexpression of human Dsg3 in several epithelial lines induces formation of membrane protrusions, a phenotype suggestive of Rho GTPase activation. Here we examined the interaction between Dsg3 and actin in detai...

Cell–cell adhesion and contraction play an essential role in the maintenance of geometric shape and polarisation of epithelial cells. However, the molecular regulation of contraction during cell elongation leading to epithelial polarisation and acquisition of geometric cell shape is not clear. Upon induction of cell–cell adhesion, we find that huma...

3-Phosphoinositide dependent protein kinase-1 (PDK1) and phospholipase C (PLC)γ1 are two key enzymes in signal transduction that control several intracellular processes. Despite the fact that PLCγ1 has been investigated for several years, the mechanisms of activation of this enzyme are still not completely clear. Similarly, although PDK1 has been m...

The neuropoietic cytokines and their cytoplasmic signaling molecules contribute to axotomy-induced events in the nerve cell body that are beneficial to axonal regeneration. Previous studies have revealed a paradox in that, in vivo, suppressor of cytokine signaling (SOCS3) is induced in axotomized primary sensory neurons which are in a growth mode b...

Control images for the AbFRET experiments, showing the level of bleed through into the FRET channel of cells labelled with the Donor alone and Acceptor alone. The FRET channel is shown using the Spectrum look up table from Image J. (9.53 MB TIF)

Expression of Dsg3 caused faster and wider spreading of 293T cells. Cells were transfected transiently with either the pBABE-GFP or -hDsg3.myc construct for two days before harvesting with trypsin. Cells then were seeded at low density onto coverslips without coating and fixed at various time points prior to immunostaining for the indicated protein...

Over-expression of Dsg3 failed to enhance cell-cell adhesion. Cells were grown to confluence before being treated with 2.4 units/ml dispase for 20 minutes to detach the epithelial sheets. The epithelial sheets were washed with PBS, twice gently, before being subjected to mechanical stress by pipetting three-five times with 1-ml blue tips. The epith...

Desmoglein 3 (Dsg3), a desmosomal adhesion protein, is expressed in basal and immediate suprabasal layers of skin and across the entire stratified squamous epithelium of oral mucosa. However, increasing evidence suggests that the role of Dsg3 may involve more than just cell-cell adhesion. To determine possible additional roles of Dsg3 during epithe...

Rho GTPases regulate multiple cellular responses, including cell motility and cell cycle progression. The Rho isoform RhoB represses transformation and affects endosomal trafficking, but its effects on cell adhesion and migration have not been investigated in detail. Here we show that RhoB-null macrophages are more rounded than wild-type macrophage...

Rac GTPases are believed to contribute to migration in leukocytes by transducing signals from cell surface receptors to the actin and microtubule cytoskeletons. Mammals have three closely related Rac isoforms, Rac1, Rac2 and Rac3, and it is widely assumed that cell migration requires the activity of these Rac GTPases. We have previously shown that...

Podosomes are actin-rich adhesive foci found in several cell types, including macrophages. They have a core containing actin and actin-binding proteins and a peripheral ring of integrins and associated proteins. We show that podosomes are abundant in polarized mouse bone marrow-derived macrophages (BMM) and are found primarily in lamellae. We inves...

Higher vertebrates have 3 Rho GTPases, RhoA, RhoB, and RhoC, which share 85% amino acid sequence identity. Here, we compare and contrast the roles of RhoA, B, and C in the regulation of the cytoskeleton and cell motility. Despite their similarity, some regulators and effectors show preferential interaction with RhoA, B, or C, and the three proteins...

The cystic fibrosis transmembrane conductance regulator (CFTR) gene shows a complex pattern of expression, with temporal and spatial regulation that is not accounted for by elements in the promoter. One approach to identifying the regulatory elements for CFTR is the mapping of DNase I hypersensitive sites (DHS) within the locus. We previously ident...