WebKEYWORDS pBABE-puro mCherry-EGFP-LC3B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6952) AUTHORS N'Diaye EN, Kajihara KK, Hsieh I, Morisaki H, Debnath J, Brown EJ TITLE PLIC proteins or ubiquilins regulate autophagy-dependent cell survival during nutrient starvation. Web3 apr. 2024 · Virus particles were collected after 48 h infection, filtered through a 0.22 μm membrane, added to HCT116 cells, and then incubated for 24 h at 37 °C. The medium was replaced and HCT116 cells were selected for puromycin tolerance (5 μg/mL) to obtain Caspase 3 knock-down, GFP-LC3B, or mcherry-EGFP-LC3B overexpression cells.

Flux-Based Assay for the Identification of Autophagy Modulators …

Web(F and G) GFP-LC3B-expressing HeLa cells were infected with mCherry-expressing Salmonellae. Correlative cryo-ET captures early phagophores (F) and expanded phagophores (G) around a damaged SCV. WebTo monitor the autophagic sequestration of p62- and LC3-positive bodies, we developed a novel pH-sensitive fluorescent tag consisting of a tandem fusion of the red, acid-insensitive mCherry and the acid-sensitive green fluorescent proteins. This approach revealed that p62- and LC3-positive bodies are degraded in autolysosomes. starbucks portable coffee mugs

Inhibition of Autophagic Degradation Process Contributes to

Webof the GFP fluorescent signal to acidic conditions of the lysosome lumen, leading to its inactivation compared with the mCherry fluorescence, which is more stable. Thus, colocalization of GFP and mCherry fluorescence (yellow puncta) indicates that the tandem protein is not localized in compartments fused with a lysosome (i.e., on the phagophore or Web13 apr. 2024 · For Airyscan confocal live-cell imaging, MatTek dishes containing infected cells producing GFP–SEPT6 and mCherry–LC3B were placed on a temperature-regulated chamber (37°C and 5% CO 2) and imaged using a 63×/1.4 C-Plan Apo oil immersion lens on a Zeiss LSM 880 confocal microscope driven by ZEN Black software. Web18 sep. 2024 · Adenovirus expressing mCherry-GFP-LC3B fusion protein (C3011), cell lysis buffer NP-40 (50 mM Tris pH 7.4, 150 mM NaCl, 1% NP-40) (P0013F) and phenylmethyl sulfonylfluoride protease inhibitor (ST505) were purchased from Beyotime (Shanghai, China). Earle’s balanced salt solution (EBSS) (14155–063) was purchased from Gibco. starbucks population health