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. Author manuscript; available in PMC: 2023 Aug 4.
Published in final edited form as: Mol Cell. 2022 Jul 28;82(15):2900–2911.e7. doi: 10.1016/j.molcel.2022.06.035

Figure 2. Unannotated microproteins and alt-proteins identified with MicroID are endogenously expressed and correctly localized to subnuclear regions of cultured human cells.

Figure 2.

(A) HEK 293T cell lines bearing epitope tag knock-ins (KI) to a genomic copy of the indicated microprotein/alt-protein coding sequence were subjected to immunofluorescence with anti-FLAG tag (magenta), colocalization marker for each indicated (sub)nuclear region (yellow, see Methods), and DAPI (cyan). Scale bar, 10 μm. Data are representative of three biological replicates. (B-E) Volcano plot of proteins enriched by co-immunoprecipitation (co-IP) from microprotein/alt-protein FLAG tag KI cells (PD, pull-down) vs. wild-type control HEK 293T nuclear lysates with label-free quantiative (LFQ) proteomics (N = 3, biological replicates). Baits and candidate interaction partner proteins known to colocalize to the same subnuclear compartment are indicated and gene names are labeled.