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To confirm that human TfR1 is present in the Affi-Gel 15-isolated proteins, including human TfR1 as known from the initial MS analyses, immunoblotting was carried out using anti-TfR1 (Abcam, Cambridge, MA, ab-84036, 1 μg/mL) and an HRP-linked second antibody (1:5,000 polyclonal rabbit anti-IgG, 65–6120, Thermo Fisher Scientific) using whole-cell extracts (15–20 µg) on 8% or 4%–20% SDS-PAGE gels, followed by transfer to PVDF membranes

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