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Localization of TfR/Cosmc/TfR (construct #5). A–D, immunofluorescent staining of TfR/Cosmc/TfR. Cells were stained with anti-HPC4 (red) antibody and anti-calnexin (green). Merge, yellow. E, sucrose gradient subcellular fractionation. The PNS was applied to a sucrose gradient and 18 fractions (top to bottom) were obtained after ultracentrifugation. Proteins from each fraction were analyzed on Western blot with anti-HPC4 and anti-KDEL antibodies.
