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Description
HeLa cells transiently expressing GlcNAcT-1-HA-KDEL were fixed, immunolabeled for GlcNAcT-1-HA-KDEL (red) and endogenous alpha-mannosidase II (ManII, green), and examined by fluorescence microscopy. Endogenous ManII displayed different staining patterns, depending upon the level of expression of GlcNAcT-1-HA-KDEL. In this image, ManII is mislocalized to the ER. Other images in Fig 6 show ManII localization to the Golgi (CIL# 13656), or cells in which ManII protein is lost completely (CIL# 13657). Transfected HeLa cells were fixed in 3% paraformaldehyde, blocked with 0.2% Triton X-100, 1% FBS, 0.5% BSA in PBS with Ca[2+] and Mg[2+], followed by incubation in primary antibodies (anti–human ManII rabbit pAb and anti-HA mouse mAb, HA.11 (Covance)) and secondary antibodies (Alexa 568 and Alexa 488) and DAPI (blue) to stain nuclei. Cells were mounted using Fluoromount (SouthernBiotech). Immunofluorescent images were digitally acquired on an inverted microscope (Axiovert 200M; Carl Zeiss, Inc.) coupled to a 12-bit cooled charge-coupled device camera (AxioCam MRm Rev. 3; Carl Zeiss, Inc.) controlled by Axiovision software (AxioVs40, version 4.7.2.0; Carl Zeiss, Inc.), using a 100x/1.3 NA oil immersion objective (EC Plan-NeoFluar; Carl Zeiss, Inc.), and saved as tiff files (1388 × 1040, 8 bit). All pictures were treated identically using Adobe Photoshop to remove background and adjust contrast. This image corresponds to Figure 6D, top panel (merge) in J Cell Biol. 190: 893-910, 2010. Images in Fig 6 include CIL#s 13655, 13656, 13657. Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp) Huang, Hung-Hsiang; Stanley, Pamela (2021). CIL:13655, Homo sapiens, epithelial cell, cervical carcinoma. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J07P8X4H
Type
image
Identifier
ark:/20775/bb6115889j
Language
No linguistic content
Subject
Protein N-linked glycosylation via asparagine HeLa Zinc ion binding Alpha-1,3-mannosylglycoprotein 2-beta-N-acetylglucosaminyltransferase activity Carbohydrate metabolic process Mannosyl-oligosaccharide 1,3-1,6-alpha-mannosidase activity Cervical carcinoma Endoplasmic reticulum-Golgi intermediate compartment Integral to membrane Endoplasmic reticulum Epithelial cell Golgi apparatus Golgi membrane Homo sapiens Cell Image Library Group ID: 6874
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