The "human cells" section gives an overview about the subcellular location of the protein of interest obtained by indirect immunofluorescence microscopy, an antibody-based protein-visualization technique. The immunofluorescent analysis is carried out in three different cell lines, one of them always being U-2 OS. A selection of immunofluorescent images is displayed below. Three different organelle probes are displayed as different channels in the multicolor images - nucleus stained in blue, microtubules in red and ER in yellow. The antibody staining targeting the protein of interest is shown in green. By using the toggle channel buttons, the different channels can be turned on and off. For the selection of the 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. For cell structure reference, visit the cell dictionary.

The siRNA validation section gives an overview of the siRNA downregulation assay performed for the antibody. For each HPA antibody, the siRNA downregulation assay is performed in duplicates with two different siRNAs. Only successful assays are included in the Atlas. Read more about siRNA validation. In the table at the top the siRNA validation score for the assay is shown together with the results from the statistical analysis of the analyzed images. The Relative Fluorescence Intensity (RFI) denotes the percentage of remaining staining intensity after siRNA downregulation. The boxplot on the top left is magnified when clicked, and illustrates the changes in fluorescent light intensity between siRNA treated and control (scrambled) samples. Also, the cell segmentation compartment, in which the changes in intensity have been measured, is noted here. siRNA transfected U-2 OS cells are immunofluorescently stained with the corresponding antibody. Images from the scrambled siRNA (negative control) well are displayed to the left and the images from one or two siRNA-assays are displayed to the right. The antibody and intensity channel toggle can be used to further illustrate the intensity downregulation when comparing images from scrambled and targeting siRNA samples in the images. 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.

For the genes where the mouse and human genes are orthologues, the mouse cell line NIH 3T3 is also stained. The main subcellular location of the encoded proteins and any additional locations are reported as well as staining characteristics, staining intensity and validation score. Example images are shown below. 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. For cell structure reference, visit the cell dictionary.