Assay for determining the subcellular location of the encoded protein(s) in human cells by indirect immunofluorescence and confocal microscopy. The antibody-based staining is generally carried out in U2OS and two additional cell lines, selected based on RNA expression. Representative multi-color images showing the protein of interest in green are displayed below. The images also include markers for the nucleus (blue), microtubules (red) and ER (yellow). All images are clickable for an enlarged view. For cell structure reference, visit the cell dictionary.

The custom data section contains information about experiments that were carried out by researchers in the Cell Atlas independent of the general workflow in the project. This includes but is not limited to colocalization experiments with known organelle markers or stainings of proteins in the cell cycle marker cell line U-2 OS FUCCI (Fluorescence Ubiquitination Cell Cycle Indicator). Example images as well as a detailed description of the respective assay are provided. Different organelle probes are displayed as different channels in the multicolor images. By using the "toggle channels"-buttons, the different channels can be turned on and off. To change which images to compare, use the checkboxes next to the images at the bottom. Three images can be compared at a time. All images are clickable for an enlarged view. The selected image will appear in large size and miniature images with all other staining results for this gene will be listed at the top left of the image. The selected miniature image has an orange overlay. If a movie is available click the thumbnail in order to show the video clip.

Assay for validating subcellular location as determined by immunofluorescence by siRNA-mediated downregulation of gene expression. The top section shows a summary of the results of the siRNA validation assay. The box plot shows the Relative Fluorescence Intensity (RFI, i.e. percentage of remaining staining intensity after siRNA downregulation) in siRNA treated and negative control (scrambled) samples. Representative images of the negative control cells are displayed to the left and representative images of siRNA-treated cells are displayed to the right. All images are clickable for an enlarged view. The selected image will appear in large size and miniature images with all other staining results for this gene will be listed at the top left of the image. The selected miniature image has an orange overlay.