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. Author manuscript; available in PMC: 2019 Sep 13.
Published in final edited form as: Nature. 2019 Mar 13;567(7749):535–539. doi: 10.1038/s41586-019-1024-7

Extended Data Figure 1. Bioorthogonal labeling by 5-PT identifies H3 serotonylation in chromatin.

Extended Data Figure 1

a, Structures of 5-HT and 5-PT in transamidation reactions. b, Immunofluorescence images (Scale bars equal 500 μm) of intracellular 5-PT in HeLa cells (bottom) after exogenous application of the molecule (vs. vehicle, top). Intracellular 5-PT was imaged in fixed cells after chemical labeling with Alexa Fluor 488 azide. DAPI was used as a nuclear co-stain. Results confirmed in ≥ 2 independent experiments. c, Cell vs. lysate donor competition assays indicating that application of excess 5-HT to live HeLa cells, but not to processed lysates, prior to chemical labeling and 5-PT-based pulldowns results in loss of H3 signal post-IP. Input and IP WBs are shown. d, Cellular fractionation analysis (WB) identifying H3 serotonylation post-IP in HeLa cell chromatin, but not in soluble nuclear or cytosolic fractions. Input and IP WBs are shown. H3 results confirmed in ≥ 3 independent experiments.