EdU Incorporation To Assess Cell Proliferation and Drug Susceptibility in Naegleria fowleri

Troth, Emma V.; Kyle, Dennis E.

doi:10.1128/aac.00017-21

Cited by 10 publications

(7 citation statements)

References 18 publications

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“…EdU is a pyrimidine analog of thymidine that can be used as a quantitative endpoint for cell proliferation by flow cytometry (Troth & Kyle, 2021). To investigate the proliferation ability of HL-60 cells or SKM-1 cells, HL-60 cells or SKM-1 cells were harvested after coculture with BM MΦs and then incubated with 50 μM EdU (RiboBio) in 48-well plates for 1 h at 37°C.…”

Section: -Ethynyl-20-deoxyuridine (Edu) Assaymentioning

confidence: 99%

Bone marrow macrophages are involved in the ineffective hematopoiesis of myelodysplastic syndromes

Xing,

Yao,

Zhao

et al. 2024

Journal Cellular Physiology

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Myelodysplastic syndromes (MDS) are a group of heterogeneous myeloid clonal disorders characterized by ineffective hematopoiesis. Accumulating evidence has shown that macrophages (MΦs) are important components in the regulation of tumor progression and hematopoietic stem cells (HSCs). However, the roles of bone marrow (BM) MΦs in regulating normal and malignant hematopoiesis in different clinical stages of MDS are largely unknown. Age‐paired patients with lower‐risk MDS (N = 15), higher‐risk MDS (N = 15), de novo acute myeloid leukemia (AML) (N = 15), and healthy donors (HDs) (N = 15) were enrolled. Flow cytometry analysis showed increased pro‐inflammatory monocyte subsets and a decreased classically activated (M1) MΦs/alternatively activated (M2) MΦs ratio in the BM of patients with higher‐risk MDS compared to lower‐risk MDS. BM MФs from patients with higher‐risk MDS and AML showed impaired phagocytosis activity but increased migration compared with lower‐risk MDS group. AML BM MΦs showed markedly higher S100A8/A9 levels than lower‐risk MDS BM MΦs. More importantly, coculture experiments suggested that the HSC supporting abilities of BM MΦs from patients with higher‐risk MDS decreased, whereas the malignant cell supporting abilities increased compared with lower‐risk MDS. Gene Ontology enrichment comparing BM MΦs from lower‐risk MDS and higher‐risk MDS for genes was involved in hematopoiesis‐ and immunity‐related pathways. Our results suggest that BM MΦs are involved in ineffective hematopoiesis in patients with MDS, which indicates that repairing aberrant BM MΦs may represent a promising therapeutic approach for patients with MDS.

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Section: -Ethynyl-20-deoxyuridine (Edu) Assaymentioning

confidence: 99%

Bone marrow macrophages are involved in the ineffective hematopoiesis of myelodysplastic syndromes

Xing,

Yao,

Zhao

et al. 2024

Journal Cellular Physiology

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“…EdU staining assay could be used to further explore the cells at the S-phase of the cell cycle to reflect real situations after the different interventions. (Okada et al, 2017;Troth & Kyle, 2021).…”

Section: Effects Of Peptide Pw5 and Rutin On D-gal Induced Oxidative ...mentioning

confidence: 99%

The study of single‐cell dynamics contributes to the evaluation of food‐derived antioxidant capacity

Xiang

Qing

et al. 2023

eFood

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Exploring potential food‐derived bioactive substances that relieve oxidative stress is considered an important goal for antioxidant research. We first used the long‐pair electron index and orbital fingerprint to predict the molecular interactions in a range of polyphenols and polypeptides. We found Rutin and peptide PW5 had the highest LPE index values. Subsequently, by D‐galactose (D‐gal)‐induced oxidative NIH3T3 cell model, we proved both Rutin and PW5 could effectively protect cells against D‐gal‐induced damage, through enhancement in cell proliferation and reduction in β‐galactosidase activity. Although Rutin displayed better performance than PW5 in the oxidative stress model, we confirmed Rutin stimulated obvious changes in the morphology and motility of normal NIH3T3 cells based on the real‐time dynamic images. Taken together, combined computational and typical model experiment methods, proved useful in efficiently screening food‐derived antioxidants. Moreover, the analytical results of cell morphology and movement may provide novel insights for the safety evaluation of antioxidants.

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“…In the EdU experiment, transfected HEC-1A cells (2×10 4 cells/well) was added to EdU medium, and fluorescence staining solution was added after incubation. The DNA replication activity of the fluorescently labeled mixture at 350 nm was tested applying a fluorescence microscope (Troth & Kyle, 2021).…”

Section: Cell Counting Kit (Cck)-8 and Edu Assaymentioning

confidence: 99%

Silencing circCAMSAP1 suppresses malignant behavior of endometrial cancer by targeting microRNA-370-3p/MAPK1

et al. 2023

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The study was conducted to figure out the function and mechanism of circular RNA circCAMSAP1 in repressing malignant behavior of endometrial carcinoma (EC) by targeting microRNA (miR)-370-3p /MAPK1. Tumor tissues and normal adjacent tissues of EC patients were harvested, and circCAMSAP1 and MAPK1 were elevated but miR-370-3p was reduced in tissues and cells of EC patients. Functional test results clarified transfection of si-circCAMSAP1 or miR-370-3p-mimic refrained cancer cell proliferation, migration and invasion, but motivated cancer cell apoptosis. Meanwhile, the amount of E-cadherin elevated and the amount of N-cadherin elevated or reduced. After co-transfection with si-circCAMSAP1 and miR-370-3p-inhibitor, miR-370-3p-inhibitor blocked si-circCAMSAP1’s therapeutic impact. Furthermore, after co-transfection of pcDNA-circCAMSAP1 and si-MAPK1, si-MAPK1 turned around the malignant effect of pcDNA-circCAMSAP1. It was testified that miR-370-3p was circCAMSAP1’s target, and inversely controlled via circCAMSAP1. Meanwhile, enhancing miR-370-3p led to repressive MAPK1, which was recognized as miR-370-3p’s downstream target. All in all, the results of this study convey silencing circCAMSAP1 refrains the malignant behavior of EC by controlling miR-370-3p /MAPK1 axis.

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EdU Incorporation To Assess Cell Proliferation and Drug Susceptibility in Naegleria fowleri

Cited by 10 publications

References 18 publications

Bone marrow macrophages are involved in the ineffective hematopoiesis of myelodysplastic syndromes

Bone marrow macrophages are involved in the ineffective hematopoiesis of myelodysplastic syndromes

The study of single‐cell dynamics contributes to the evaluation of food‐derived antioxidant capacity

Silencing circCAMSAP1 suppresses malignant behavior of endometrial cancer by targeting microRNA-370-3p/MAPK1

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