siRNA and pharmacological inhibition of endocytic pathways to characterize the differential role of macropinocytosis and the actin cytoskeleton on cellular uptake of dextran and cationic cell penetrating peptides octaarginine (R8) and HIV-Tat

Soraj, Monerah Al; He, Lin; Peynshaert, Karen; Cousaert, Julie; Vercauteren, Dries; Braeckmans, Kevin; Smedt, Stefaan C. De; Jones, Arwyn Tomos

doi:10.1016/j.jconrel.2012.03.015

Cited by 93 publications

(114 citation statements)

References 56 publications

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“…To determine which pathways JosD1 affects, we examined the uptake of fluorescence markers specific for these three pathways: Dex-TMR (macropinocytosis), Tf-Alexa Fluor 568 (clathrin-mediated endocytosis), and CTxB-Alexa Fluor 555 (caveolae-mediated endocytosis) (21,22).…”

Section: Resultsmentioning

confidence: 99%

JosD1, a Membrane-targeted Deubiquitinating Enzyme, Is Activated by Ubiquitination and Regulates Membrane Dynamics, Cell Motility, and Endocytosis

Seki

Gong

Williams

et al. 2013

Journal of Biological Chemistry

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Background:The functions of the deubiquitinating enzymes JosD1 and JosD2, related to ATXN3, are unknown. Results: JosD1 is activated by ubiquitination, localizes to plasma membrane, and affects membrane dynamics, cell motility, and endocytosis. Conclusion: JosD1 and JosD2 possess divergent properties, with JosD1 regulating membrane-related functions. Significance: Our findings provide insight into diverse functions of a disease-linked family of deubiquitinating enzymes.

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Section: Resultsmentioning

confidence: 99%

JosD1, a Membrane-targeted Deubiquitinating Enzyme, Is Activated by Ubiquitination and Regulates Membrane Dynamics, Cell Motility, and Endocytosis

Seki

Gong

Williams

et al. 2013

Journal of Biological Chemistry

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“…After exposure the cells were washed twice in cold washing buffer (Hepes 10 mM, NaN 3 5 mM) and the remaining membrane bound label was removed according to previously published methods [12, 13]. Briefly, 1 min incubation in ice-cold acidic wash (0.2 M acetic acid, 0.2 M NaCl, pH 2.0), followed by two washes in cold washing buffer was employed to remove surface-bound Transferrin.…”

Section: Methodsmentioning

confidence: 99%

The agglomeration state of nanoparticles can influence the mechanism of their cellular internalisation

et al. 2017

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BackgroundSignificant progress of nanotechnology, including in particular biomedical and pharmaceutical applications, has resulted in a high number of studies describing the biological effects of nanomaterials. Moreover, a determination of so-called “critical quality attributes”, that is specific physicochemical properties of nanomaterials triggering the observed biological response, has been recognised as crucial for the evaluation and design of novel safe and efficacious therapeutics. In the context of in vitro studies, a thorough physicochemical characterisation of nanoparticles (NPs), also in the biological medium, is necessary to allow a correlation with a cellular response. Following this concept, we examined whether the main and frequently reported characteristics of NPs such as size and the agglomeration state can influence the level and the mechanism of NP cellular internalization.ResultsWe employed fluorescently-labelled 30 and 80 nm silicon dioxide NPs, both in agglomerated and non-agglomerated form. Using flow cytometry, transmission electron microscopy, the inhibitors of endocytosis and gene silencing we determined the most probable routes of cellular uptake for each form of tested silica NPs. We observed differences in cellular uptake depending on the size and the agglomeration state of NPs. Caveolae-mediated endocytosis was implicated particularly in the internalisation of well dispersed silica NPs but with an increase of the agglomeration state of NPs a combination of endocytic pathways with a predominant role of macropinocytosis was noted.ConclusionsWe demonstrated that the agglomeration state of NPs is an important factor influencing the level of cell uptake and the mechanism of endocytosis of silica NPs.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-017-0281-6) contains supplementary material, which is available to authorized users.

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“…Further, the distribution pattern of the AMPs in HeLa WT cells was assayed by confocal microscopy with and without fluorophore-labeled dextran as an endocytic probe that can be used to label the entire endolysosomal pathway or specifically lysosomes (28), and which under the applied experimental conditions accumulate in the late vacuolar system, particularly in the lysosomes. Fluorophore-labeled transferrin known to be taken up by clathrin-dependent endocytosis (29) was included to investigate a potential contribution to the uptake via this pathway. Expectedly, penetratin showed a punctate uptake pattern with some co-localization with dextran, and no or poor co-localization with transferrin indicating that its uptake is not receptor-mediated.…”

Section: Discussionmentioning

confidence: 99%

Cell-Penetrating Antimicrobial Peptides – Prospectives for Targeting Intracellular Infections

et al. 2015

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siRNA and pharmacological inhibition of endocytic pathways to characterize the differential role of macropinocytosis and the actin cytoskeleton on cellular uptake of dextran and cationic cell penetrating peptides octaarginine (R8) and HIV-Tat

Cited by 93 publications

References 56 publications

JosD1, a Membrane-targeted Deubiquitinating Enzyme, Is Activated by Ubiquitination and Regulates Membrane Dynamics, Cell Motility, and Endocytosis

JosD1, a Membrane-targeted Deubiquitinating Enzyme, Is Activated by Ubiquitination and Regulates Membrane Dynamics, Cell Motility, and Endocytosis

The agglomeration state of nanoparticles can influence the mechanism of their cellular internalisation

Cell-Penetrating Antimicrobial Peptides – Prospectives for Targeting Intracellular Infections

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