Fig. 1: Stability of the assembled nuclear pore upon rapid loss of BSK-NUPs.

a Schematic of CRISPR/Cas9-based tagging of nucleoporin genes and subsequent degradation of the AID-fused proteins. Each AID cell line is biallelically tagged. b NUP98, NUP133, and RANBP2 localization is not altered upon loss of NUP153 or TPR (4 h of auxin treatment). AID-NG tagged NUP153 and TPR proteins are shown in green, antibody-stained NUPs in red. Data are representative of at least three independent experiments. c Heat maps of differential abundance of NUP153, NUP50, and TPR proteins in NPC-enriched nuclear extracts and total cell lysates of the indicated AID-tagged cells 2 h after auxin treatment using TMT-assisted and conventional (label-free) mass spectrometry. Green color indicates a decrease in protein abundance after auxin treatment, log2 scale. Data represent a repeat of one experiment. d Localization of BSK-NUPs in the absence or presence of TPR and NUP153 (4 h of auxin treatment). Plots show line scans of relative fluorescence intensity (RFU) for BSK-NUPs (AID-NG tagged NUPs are shown in green, antibody-stained NUPs in red) and DNA across the nucleus. DNA was counterstained with DAPI (gray). Note that NUP50 no longer localizes to the nuclear envelope (NE) in the absence of NUP153. Scale bar: 5 µm. Data are representative of at least three independent experiments. AID auxin-inducible degron, TMT tandem mass tag, FC fold change.