Autophagic flux assay chloroquine

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  1. lopster User

    Autophagic flux assay chloroquine


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    Annexin V–FITC/PI double-staining assay, the activity and activating cleavages of caspase-3, -8, and -9. MTT assay and colony forming assay were used to assess the effect of matrine on growth and proliferation of HCC cells. Autophagic flux in HCC cells was analyzed using the expression of LC3BI/II and p62/SQSTM1, GFP-LC3 transfection, and First, we performed an in vivo autophagic flux assay by treating the rats 48 hours before the end of the experimental protocol with colchicine 0.4 −1, a microtubule depolarizing agent. Although this assay can be used as an indicator of autophagic flux, it does not provide precisely the same information as other flux assays that directly measure endpoints of lysosomal degradation. This method depends on the acidification and degradation capacity of the lysosome.

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    Autophagic flux assay chloroquine

    Inhibition of autophagic flux differently modulates., Exercise prevents impaired autophagy and proteostasis in a model of.

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  7. CYTO-ID ® Autophagy Detection Kit measures autophagic vacuoles and monitors autophagic flux in lysosomally inhibited live cells using a novel dye that selectively labels accumulated autophagic vacuoles. The dye has been optimized through the identification of titratable functional moieties that allow for minimal staining of lysosomes while.

    • CYTO-ID® Autophagy detection kit - ENZ-51031 - Enzo Life Sciences.
    • Methods in Mammalian Autophagy Research - PubMed Central PMC.
    • Autophagy Assay LC3 Autophagic Flux Assay Promega.

    Methyl adenine 3-MA, wortmannin, bafilomycin A1, chloroquine, NH4Cl, or leupeptin, to block lysosomal degradation and determine the non-autophagic protein degradation background 12 Table2. The detailed methodology for the execution of the long-lived protein degradation assay is explained in detail in the literature 12. Table 2. Transmission electron microscopy, mRFP-GFP-LC3 assay and p62 protein degradation, confirmed that Luteolin stimulated the autophagic process in the metastatic MET4 cells. Blocking autophagy using chloroquine magnified Luteolin-induced apoptosis in the metastatic SCC cells. A549 cells treated to induce autophagy and imaged. Left A549 cells were treated with chloroquine and stained with Invitrogen Hoechst 33342, Invitrogen HCS CellMask Deep Red, and an anti-LC3B with Invitrogen Alexa Fluor 488 goat anti-rabbit antibody.

     
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