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Addgene inc rfp lc3
Rfp Lc3, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pmrfp+lc3/pmRFP-LC3+(Plasmid+%2321075)/pm41797117-56-4-13
Average 93 stars, based on 140 article reviews
rfp lc3 - by Bioz Stars, 2026-09
93/100 stars

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Article Title: STX1A localizes to the lysosome and controls its exocytosis
Article Snippet: Lysosome exocytosis is one of the critical functions of lysosomes in maintaining cellular homeostasis and plasma membrane repair.. At the basal level, the SNAREs regulating the lysosome fusion with the cell surface have been poorly defined.. Here, we identified a Qa-SNARE STX1A, localized majorly to lysosomes and a cohort to the plasma membrane in HeLa cells.

Article Title: STX1A localizes to the lysosome and controls its exocytosis
Article Snippet: Other plasmids: mCherry-UtrCh (26740), GFP-CD63 (62964), LAMP1-GFP (34831), LAMP1-RFP (1817), pMD2.G (VSV-G lentiviral envelop vector, 12259), psPAX2 (lentiviral packaging vector, 12260), pmRFP-LC3 (21075) and ptLC3 (rat LC3 fused to mRFP and EGFP, 21074) were obtained from Addgene.

Transfection:

Article Title: Endosome-phagophore linking assemblies for the degradation of membrane/extracellular proteins.
Article Snippet: .. HEK293T cells were transfected with the 3:1 (mass ratio) complexes of PEI and plasmids including pEGFP-N1/PD-L1, GFP-Rab5B (addgene Cat# 61802), pmRFP-LC3 (addgene Cat# 21075), and mTagBFP2-Lysosomes-20 (addgene Cat# 55308) in serum-free DMEM for 24 h. The transfected cells were further treated with 0.5 μM different assemblies for 12 h or incubated with 0.5 μM CVR-EPLA for various times, then fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope. ..

Article Title: HOPS-dependent vesicle tethering lock inhibits endolysosomal fusions and autophagosome secretion upon the loss of Syntaxin17
Article Snippet: .. We used pmRFP-LC3 (21075; deposited by T. Yoshimori—purchased from Addgene) and LAMP1-mGFP plasmids (34831; deposited by E. Dell’Angelica—purchased from Addgene) for transient transfection ( ). .. This round of transfection was carried out at around 80% confluency using TransIT-LT1 transfection reagent (Mirus 22,084,721) with the desired plasmids following the manufacturer’s instructions using 1 to 2 μg each of the plasmid DNA.

Article Title: Endosome-phagophore linking assemblies for the degradation of membrane/extracellular proteins
Article Snippet: HEK293T cells were transfected with the 3:1 (mass ratio) complexes of polyethylenimine (PEI, 1 mg mL −1 and pH 6.9–7.1, Kyfora Bio Cat# 23966) and plasmids including pEGFP-N1/PD-L1 (addgene Cat# 121478) and BFP-Rab5 (addgene Cat# 49147) in serum-free DMEM for 24 h. The transfected cells were further incubated with 0.5 μM CVR-EPLA for various times, fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope (NIS Viewer 5.21 and NIS-Elements Viewer 5.21.00, Nikon, Japan). .. HEK293T cells were transfected with the 3:1 (mass ratio) complexes of PEI and plasmids, including pEGFP-N1/PD-L1, GFP-Rab5B (addgene Cat# 61802), pmRFP-LC3 (addgene Cat# 21075), and mTagBFP2-Lysosomes-20 (addgene Cat# 55308) in serum-free DMEM for 24 h. The transfected cells were further treated with 0.5 μM different assemblies for 12 h or incubated with 0.5 μM CVR-EPLA for various times, then fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope. ..

Article Title: HOPS-dependent vesicle tethering lock inhibits endolysosomal fusions and autophagosome secretion upon the loss of Syntaxin17.
Article Snippet: .. We used pmRFP- LC3 (21075; deposited by T. Yoshimori—purchased from Addgene) and LAMP1- mGFP plasmids (34831; deposited by E. Dell’Angelica—purchased from Addgene) for transient transfection (16). .. This round of transfection was carried out at around 80% confluency using TransIT- LT1 transfection reagent (Mirus 22,084,721) with the desired plasmids following the manufacturer’s instructions using 1 to 2 μg each of the plasmid DNA.

Incubation:

Article Title: Endosome-phagophore linking assemblies for the degradation of membrane/extracellular proteins.
Article Snippet: .. HEK293T cells were transfected with the 3:1 (mass ratio) complexes of PEI and plasmids including pEGFP-N1/PD-L1, GFP-Rab5B (addgene Cat# 61802), pmRFP-LC3 (addgene Cat# 21075), and mTagBFP2-Lysosomes-20 (addgene Cat# 55308) in serum-free DMEM for 24 h. The transfected cells were further treated with 0.5 μM different assemblies for 12 h or incubated with 0.5 μM CVR-EPLA for various times, then fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope. ..

Article Title: Endosome-phagophore linking assemblies for the degradation of membrane/extracellular proteins
Article Snippet: HEK293T cells were transfected with the 3:1 (mass ratio) complexes of polyethylenimine (PEI, 1 mg mL −1 and pH 6.9–7.1, Kyfora Bio Cat# 23966) and plasmids including pEGFP-N1/PD-L1 (addgene Cat# 121478) and BFP-Rab5 (addgene Cat# 49147) in serum-free DMEM for 24 h. The transfected cells were further incubated with 0.5 μM CVR-EPLA for various times, fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope (NIS Viewer 5.21 and NIS-Elements Viewer 5.21.00, Nikon, Japan). .. HEK293T cells were transfected with the 3:1 (mass ratio) complexes of PEI and plasmids, including pEGFP-N1/PD-L1, GFP-Rab5B (addgene Cat# 61802), pmRFP-LC3 (addgene Cat# 21075), and mTagBFP2-Lysosomes-20 (addgene Cat# 55308) in serum-free DMEM for 24 h. The transfected cells were further treated with 0.5 μM different assemblies for 12 h or incubated with 0.5 μM CVR-EPLA for various times, then fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope. ..

Laser-Scanning Microscopy:

Article Title: Endosome-phagophore linking assemblies for the degradation of membrane/extracellular proteins.
Article Snippet: .. HEK293T cells were transfected with the 3:1 (mass ratio) complexes of PEI and plasmids including pEGFP-N1/PD-L1, GFP-Rab5B (addgene Cat# 61802), pmRFP-LC3 (addgene Cat# 21075), and mTagBFP2-Lysosomes-20 (addgene Cat# 55308) in serum-free DMEM for 24 h. The transfected cells were further treated with 0.5 μM different assemblies for 12 h or incubated with 0.5 μM CVR-EPLA for various times, then fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope. ..

Article Title: Endosome-phagophore linking assemblies for the degradation of membrane/extracellular proteins
Article Snippet: HEK293T cells were transfected with the 3:1 (mass ratio) complexes of polyethylenimine (PEI, 1 mg mL −1 and pH 6.9–7.1, Kyfora Bio Cat# 23966) and plasmids including pEGFP-N1/PD-L1 (addgene Cat# 121478) and BFP-Rab5 (addgene Cat# 49147) in serum-free DMEM for 24 h. The transfected cells were further incubated with 0.5 μM CVR-EPLA for various times, fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope (NIS Viewer 5.21 and NIS-Elements Viewer 5.21.00, Nikon, Japan). .. HEK293T cells were transfected with the 3:1 (mass ratio) complexes of PEI and plasmids, including pEGFP-N1/PD-L1, GFP-Rab5B (addgene Cat# 61802), pmRFP-LC3 (addgene Cat# 21075), and mTagBFP2-Lysosomes-20 (addgene Cat# 55308) in serum-free DMEM for 24 h. The transfected cells were further treated with 0.5 μM different assemblies for 12 h or incubated with 0.5 μM CVR-EPLA for various times, then fixed with a 4% PFA solution for 15 min, and finally imaged using a confocal laser scanning microscope. ..

Plasmid Preparation:

Article Title: Presenilins as hub proteins controlling the endocytic and autophagic pathways and small extracellular vesicle secretion
Article Snippet: .. We thank Qing Zhong for EGFP‐RAB5A‐Q79L (Addgene plasmid # 28046; http://n2t.net/addgene:28046; RRID:Addgene_28046) (Sun et al., ) and pmRFP‐LC3 from Tamotsu Yoshimori (Addgene plasmid # 21075; http://n2t.net/addgene: 21075; RRID:Addgene_21075) (Kimura et al., ). .. Mouse embryonic fibroblasts (MEFs) (wildtype (WT), PSEN1 −/− /PSEN2 −/− (PSEN‐dKO) and PSEN1 −/− (PSEN1‐KO) (Herreman et al., ) (kind gift from Dr. De Strooper), as well as WT and PSEN‐dKO (kind gift from Dr. Thinakaran), the latter into which human PSEN1WT (PSEN1c) or deltaE9‐PSEN1 (PSEN1dE9c) (Ratovitski et al., ) were stably expressed, were cultured in Dulbecco's modified Eagle's medium supplemented with 10% foetal calf serum (FCS), penicillin (100 U/mL) and streptomycin (50 μg/mL) (Thermo Fisher scientific, France) at 37°C/5% CO 2 .



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