Rotimi Fasimoye
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View article: Tagless LysoIP for immunoaffinity enrichment of native lysosomes from clinical samples
Tagless LysoIP for immunoaffinity enrichment of native lysosomes from clinical samples Open
Lysosomes are implicated in a wide spectrum of human diseases, including monogenic lysosomal storage disorders (LSDs), age-associated neurodegeneration, and cancer. Profiling lysosomal content using tag-based lysosomal immunoprecipitation …
View article: Targeted dephosphorylation of SMAD3 as an approach to impede TGF-β signaling
Targeted dephosphorylation of SMAD3 as an approach to impede TGF-β signaling Open
TGF-β (transforming growth factor-β) signaling is involved in a myriad of cellular processes and its dysregulation has been implicated in many human diseases, including fibrosis and cancer. TGF-β transcriptional responses are controlled by…
View article: Tagless LysoIP method for molecular profiling of lysosomal content in clinical samples
Tagless LysoIP method for molecular profiling of lysosomal content in clinical samples Open
Lysosomes are implicated in a wide spectrum of human diseases including monogenic lysosomal storage disorders (LSDs), age-associated neurodegeneration and cancer. Profiling lysosomal content using tag-based lysosomal immunoprecipitation (L…
View article: A nematode-specific ribonucleoprotein complex mediates interactions between the major nematode spliced leader snRNP and its target pre-mRNAs
A nematode-specific ribonucleoprotein complex mediates interactions between the major nematode spliced leader snRNP and its target pre-mRNAs Open
Spliced leader trans-splicing of pre-mRNAs is a critical step in the gene expression of many eukaryotes. How the spliced leader RNA and its target transcripts are brought together to form the trans-spliceosome remains an important unanswer…
View article: Colocalisation imaging of endogenous TMEM192 with lysosomal and mitochondria markers v1
Colocalisation imaging of endogenous TMEM192 with lysosomal and mitochondria markers v1 Open
Immunofluorescent (IF) microscopy is a powerful tool used in cellular and molecular biology to monitor the subcellular localisation of proteins. By combining the advantages of immunostaining and confocal light microscope, IF microscopy can…
View article: Cathepsin D assay to verify the retention of lysosomal content v1
Cathepsin D assay to verify the retention of lysosomal content v1 Open
Cathepsin D assay is a fluorescence-based assay that leverage on the activity of cathepsin D, a lysosomal enzyme, to monitor the intactness of lysosome in the cell. Here, we describe a method where we used the measurement of cathepsin D ac…
View article: Genetic Associations and Functional Pathways in Parkinson's Disease: A Meta-Analytic Insight into LRRK2 Mutation Impact
Genetic Associations and Functional Pathways in Parkinson's Disease: A Meta-Analytic Insight into LRRK2 Mutation Impact Open
Background: Parkinson's Disease (PD) is a complex neurodegenerative disorder with elusive pathophysiological mechanisms. Recent studies have highlighted the role of genetic variations and metabolic changes in PD progression. Aim: This stud…
View article: Parkinson’s VPS35[D620N] mutation induces LRRK2-mediated lysosomal association of RILPL1 and TMEM55B
Parkinson’s VPS35[D620N] mutation induces LRRK2-mediated lysosomal association of RILPL1 and TMEM55B Open
We demonstrate that the Parkinson’s VPS35[D620N] mutation alters the expression of ~220 lysosomal proteins and stimulates recruitment and phosphorylation of Rab proteins at the lysosome. This recruits the phospho-Rab effector protein RILPL…
View article: Introducing GolgiTAG to Cells and Immunoprecipitation of Golgi v2
Introducing GolgiTAG to Cells and Immunoprecipitation of Golgi v2 Open
Golgi apparatus is essential to the secretory pathway of the cell and by extension helps to maintain cellular homeostasis. It is the main transportation hub of the cell, where molecules destined for other organelles within the cytoplasm or…
View article: Transfection by electroporation of GFP-LRRK2 and Immunofluorescent imaging of MEFs VPS35 (D620N) mutants stably expressing LysoTag v1
Transfection by electroporation of GFP-LRRK2 and Immunofluorescent imaging of MEFs VPS35 (D620N) mutants stably expressing LysoTag v1 Open
Transfection of foreign DNA and Immunofluorescent (IF) microscopy are powerful tools used in cellular and molecular biology to monitor subcellular localisation of proteins. Mouse embryonic fibroblasts (MEFs) are primary cells notorious for…
View article: Golgi-IP, a tool for multimodal analysis of Golgi molecular content
Golgi-IP, a tool for multimodal analysis of Golgi molecular content Open
The Golgi is a membrane-bound organelle that is essential for protein and lipid biosynthesis. It represents a central trafficking hub that sorts proteins and lipids to various destinations or for secretion from the cell. The Golgi has emer…
View article: Organelle isolation from mouse tissues expressing organelle tags v1
Organelle isolation from mouse tissues expressing organelle tags v1 Open
We describe here a method to perform the rapid isolation of intact lysosomes from mouse tissues expressing a lysosome-localized TMEM192-3×HA fusion protein (“LysoTag”) (as in JAX stock #035401, RRID:IMSR_JAX:035401). The fusion protein mar…
View article: Parkinson's VPS35[D620N] retromer lysosomal dysfunction induces LRRK2 mediated lysosomal association of RILPL1 and TMEM55B
Parkinson's VPS35[D620N] retromer lysosomal dysfunction induces LRRK2 mediated lysosomal association of RILPL1 and TMEM55B Open
Primary data associated with the manuscript "Parkinson’s VPS35[D620N] mutation induces LRRK2 mediated lysosomal association of RILPL1 and TMEM55B" (Prosenjit Pal, Matthew Taylor, Pui Yiu Lam, Francesca Tonelli, Chloe A.Hecht, Pawel Lis, Ra…
View article: Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content
Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content Open
The Golgi is a membrane-bound organelle that is essential for protein and lipid biosynthesis. It represents a central trafficking hub that sorts proteins and lipids to various destinations or for secretion from the cell. The Golgi has emer…
View article: Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content
Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content Open
Raw data files used for the manuscript "Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content". The Word document (GolgiTAG paper_ file description Zenodo.docx) contains a description of each file.
View article: Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content
Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content Open
Raw data files used for the manuscript "Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content". The Word document (GolgiTAG paper_ file description Zenodo.docx) contains a description of each file.
View article: Preparation and imaging of enriched Golgi from GolgiTAG-IP using Transmission Electron Microscopy v1
Preparation and imaging of enriched Golgi from GolgiTAG-IP using Transmission Electron Microscopy v1 Open
Transmission electron microscopy (TEM) is a tool used to image, in good resolution, the structure of organelles within the cell. Available protocols are designed to image structures within fixed intact cells. Here, we described a protocol …
View article: Immunofluorescence imaging of cells expressing GolgiTAG (TMEM115-3HA) or HA-tagged non-Golgi annotated proteins v1
Immunofluorescence imaging of cells expressing GolgiTAG (TMEM115-3HA) or HA-tagged non-Golgi annotated proteins v1 Open
Immunofluorescent (IF) microscopy is a powerful tool used in cellular and molecular biology to monitor the subcellular localisation of proteins. By combining the advantages of immunostaining and confocal light microscope, IF microscopy can…
View article: Generation of stable cell lines via retroviral or lentiviral transduction v1
Generation of stable cell lines via retroviral or lentiviral transduction v1 Open
The ability to stably express a protein of interest in cells is critical to study its function. Here, we describe a protocol to generate stable cell lines using a retrovirus system that can be used for a variety of mouse and human cell lin…
View article: Immunoblotting analysis of samples from GolgiTAG (TMEM115-3HA) immunoprecipitation v1
Immunoblotting analysis of samples from GolgiTAG (TMEM115-3HA) immunoprecipitation v1 Open
Analysis of the expression of organelle-specific markers is essential to verify the efficiency of any Golgi immunoprecipitation (IP) protocol. Here, we describe our immunoblotting method to assess efficient enrichment of Golgi proteins in …
View article: A novel, essential<i>trans</i>-splicing protein connects the nematode SL1 snRNP to the CBC-ARS2 complex
A novel, essential<i>trans</i>-splicing protein connects the nematode SL1 snRNP to the CBC-ARS2 complex Open
Spliced leader trans-splicing is essential for gene expression in many eukaryotes. To elucidate the molecular mechanism of this process, we characterise the molecules associated with the Caenorhabditis elegans major spliced leader snRNP (S…
View article: Development of BromoTag: A “Bump-and-Hole”–PROTAC System to Induce Potent, Rapid, and Selective Degradation of Tagged Target Proteins
Development of BromoTag: A “Bump-and-Hole”–PROTAC System to Induce Potent, Rapid, and Selective Degradation of Tagged Target Proteins Open
Small-molecule-induced protein depletion technologies, also called inducible degrons, allow degradation of genetically engineered target proteins within cells and animals. Here, we design and develop the BromoTag, a new inducible degron sy…
View article: Introducing GolgiTAG to Cells and Immunoprecipitation of Golgi v1
Introducing GolgiTAG to Cells and Immunoprecipitation of Golgi v1 Open
Golgi apparatus is essential to the secretory pathway of the cell and by extension helps to maintain cellular homeostasis. It is the main transportation hub of the cell, where molecules destined for other organelles within the cytoplasm or…
View article: In Silico Analysis of Interaction between Seaweed-Derived Bioactive Compounds and Selected Diabetes-Related Targets
In Silico Analysis of Interaction between Seaweed-Derived Bioactive Compounds and Selected Diabetes-Related Targets Open
Seaweeds are known for their beneficial health effects in the management of diabetes mellitus (DM). Numerous bioactive metabolites of diverse chemical structures have been found in the marine algae with attributed potent pharmacological ef…
View article: An in vivo genetic screen for genes involved in spliced leader trans-splicing indicates a crucial role for continuous de novo spliced leader RNP assembly
An in vivo genetic screen for genes involved in spliced leader trans-splicing indicates a crucial role for continuous de novo spliced leader RNP assembly Open
Spliced leader (SL) trans-splicing is a critical element of gene expression in a number of eukaryotic groups. This process is arguably best understood in nematodes, where biochemical and molecular studies in Caenorhabditis elegans and Asca…
View article: PHYTOCHEMICAL EVALUATION AND IN VITRO ANTIOXIDANT STATUS OF CLERODENDRUM VOLUBILE (AN INDIGENOUS MEDICINAL PLANT)
PHYTOCHEMICAL EVALUATION AND IN VITRO ANTIOXIDANT STATUS OF CLERODENDRUM VOLUBILE (AN INDIGENOUS MEDICINAL PLANT) Open
Clerodenrum volubile is an important locally used medicinal plant. It is one of the essential herbs nature has provided for mankind to be consumed as spices, vegetable and also used in the treatment of diseases. Qualitative phytoconstituen…