Puncta formation assay

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August undefined, 2024

WebLC3 is a marker protein that is involved in the formation of autophagosomes and ... FP-LC3 and even endogenous LC3 can also be incorporated into intracellular pro … LC3 … WebJul 24, 2007 · GFP-WIPI-1 puncta-formation assay investigating different autophagy modulating agentsInduction of autophagy by prominent inducers such as rapamycin, amino acid deprivation, gleevec [16] and thapsigargin [17] was apparent employing the GFP-WIPI …

Sestrin2-mediated disassembly of stress granules dampens …

WebJun 28, 2024 · To exclude the possibility that the formation of IRS-1 puncta was due to high levels of overexpressed protein, we first established an IRS-1 knockout (KO) C2C12 cell … WebTaken together, these data indicate that galectin puncta formation can be used as a rapid and specific assay for LMP in various experimental settings, including high-throughput ... megan austin facebook

The mCherry-GFP-LC3B puncta formation assays. Confocal …

WebJun 28, 2024 · To exclude the possibility that the formation of IRS-1 puncta was due to high levels of overexpressed protein, we first established an IRS-1 knockout (KO) C2C12 cell line (C2C12-IRS-1 KO ... WebApr 11, 2024 · Cell counting kit-8 (CCK8) (Vazyme, A311-01) assay was performed to examine the cytotoxicity of rapamycin or 3-MA to RD cells. Western blotting. Western blotting was performed as described previously . ... In cells transfected with GFP-LC3, the level of LC3 puncta formation was increased by echovirus infection (Figures 3A, B). megan autrey lasting freedom llc

Resveratrol, an activator of SIRT1, induces protective autophagy in …

Quantifying autophagy: Measuring LC3 puncta and autolysosome …

WebApr 10, 2024 · Glycyrrhizin treatment for 24 h at different concentrations (50, 100, 200 μM) did not show significant toxicity. Further, we confirmed glycyrrhizin-induced autophagy by immunofluorescence of LC3B. Drug treatment for 4 h showed a significant enhancement of LC3B puncta formation by 3.2 fold change (unpaired t-test and p value: 0.0071) (Fig. 1B WebSep 22, 2024 · 3.4.2 Puncta Formation Assay. 1. Prior to experiment, thaw RAW 264.7 cells expressing GFP-LC3 and culture them. 2. When the confluence of cells becomes about 80%, harvest the cells and seed on sterile coverslips (18 × 18 cm 2) placed in 12-well plate and keep overnight at 37 °C in CO 2 incubator for adherence (see Note 11). 3. megan auto worksWebAug 31, 2012 · Both RNAi lines for Atg16 show a similar block of Lysotracker puncta formation in L3 (panels A and C), while the size of these dots is only reduced significantly by Atg16 HMS in L2 (compare D to B; see also panel E for statistics). * indicates a significant difference (p<0.05), ** indicates a very significant difference (p<0.01), based on two ... nam khong westerly ri

"WebMar 30, 2024 · For colony formation assays, cells were plated into 6-well culture plates at 1000–3000 cells/well, cultured for 14 days before being fixed with 4% paraformaldehyde ... A–D The impact of ASS1 knockdown on the formation of LC3B puncta in KYSE30 cells (A) and KYSE450 cells (B) with PSMD2 overexpression. " - Puncta formation assay

Puncta formation assay

Methods to Detect Loss of Lysosomal Membrane Integrity

WebJun 27, 2007 · GFP-WIPI-1 puncta-formation assay investigating different autophagy modulating agents Induction of autophagy by prominent inducers such as rapamycin, … Web3.3. GFP-WIPI-1 puncta-formation assay investigating diﬀerent autophagy modulating agents Induction of autophagy by prominent inducers such as rap-amycin, amino acid deprivation, gleevec [16] and thapsigargin [17] was apparent employing the GFP-WIPI-1 puncta-forma-tion assay in HeLa cells (Fig. 4a). WIPI-1 puncta/non-puncta

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WebNational Center for Biotechnology Information WebJan 5, 2024 · Galectin puncta formation assay with co-staining for lysosomal markers in fixed cells. MCF7 cells were treated with 2 mM LLOMe for 2 h, fixed and co-stained with …

WebJun 6, 2016 · During autophagy, cytoplasmic LC3 is processed and recruited to the autophagosomal membranes; the autophagosome then fuses with the lysosome to form … WebSep 22, 2024 · 3.4.2 Puncta Formation Assay. 1. Prior to experiment, thaw RAW 264.7 cells expressing GFP-LC3 and culture them. 2. When the confluence of cells becomes about …

WebApr 13, 2024 · Introdução: Casos graves de COVID-19 são caracterizados por um forte processo inflamatório que pode levar à falência de órgãos e morte do paciente. O inflamassoma de NLRP3 é uma plataforma molecular que... WebThis can also be confirmed by puncta formation assay using confocal microscopy. Further, the autophagic flux can be examined using bafilomycin A1 as inhibitor of autophagosome …

WebJan 1, 2009 · Figure 16.1. Schematic demonstration of the WIPI‐1 puncta‐formation assay ( Proikas‐Cezanne et al., 2007) 3. Indirect Immunofluorescence to Visualize Endogenous …

WebDownload scientific diagram The mCherry-GFP-LC3B puncta formation assays. Confocal images showed distribution of LC3B during the process of autophagy in HepG2 cells (A) … megan auer longmont clinicWebFeb 5, 2024 · Puncta formation was reversible, ... assay, which is based on formation of a fluorescent complex when two proteins fused to non-fluorescent fragments of a fluorescent protein interact with each ... meganatural®-bp grape seed extractWebMay 8, 2024 · e Puncta formation summary for co-expression of GFP-Par3 WT, ... both the puncta formation assay in COS7 cells and the rescue assay in fly NBs were … nam king chinese memphisWebCurrently, the best method for detecting LMP is the galectin puncta formation assay which can be used for a broad range of sample types, both fixed and live, is easy to perform, and … megan auto works villa park ilWebDec 1, 2024 · In fact, some of the autophagy activators identified through the use of the LC3 puncta formation assay later turned out to be autophagy inhibitors at downstream steps [22]. While cell-based autophagy screens have the advantage of discovering cell-active chemical probes to aid basic research, rigorous downstream work is therefore needed to … megan austin ohio universityWebJun 2, 2011 · The most popular of the assays using LC3 are microscopy-based green fluorescent protein (GFP)-LC3 puncta formation assays and western blots of LC3-I and … namkhong guesthouseWebDec 8, 2024 · GFP-LC3 puncta formation assay showed that the marked accumulation of GFP-LC3 puncta upon glucose starvation, when treated with Bafilomycin A1, was attenuated in Pontin RA MEFs compared to Pontin ... megan avenue 1 directory