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. 2022 Jun 20;24(7):1038–1048. doi: 10.1038/s41556-022-00931-x

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 2 — a. Titration of monoclonal human GPRC5C antibody. HEK293T transfected with overexpressing GPRC5C or control plasmid and stained with decreasing concentration (left to right) of the GPRC5C antibody. b. Representative FACS scheme for gating of GPRC5C^pos/neg-HSCs. GPRC5C^pos cells are identified using a fluorescence minus one (FMO) or IgG control. c. qPCR validation of GPRC5C RNA expression from BM GPRC5C^pos/neg-HSCs. Normalized to housekeeping gene GAPDH and GPRC5C^neg. n = 6 biological replicates. d. Representative FACS plots of cell cycle profiles from BM GPRC5C^pos/neg-HSCs. e. qPCR of CDK6 expression from GPRC5C^pos/neg-HSCs. Normalized to GAPDH gene and then GPRC5C^neg-HSCs. n = 8 biological replicates. f. FACS measurement of CDK6 MFI in BM GPRC5C^pos/neg-HSCs. n = 6 biological replicates. g. Representative FACS histogram of CellROX Deep Red MFI in BM GPRC5C^pos/neg-HSCs. h. t-SNE representation of the clusters from BM GPRC5C^pos/neg-HSCs. i. t-SNE depicting BM GPRC5C^pos/neg-HSCs. j. Projection of the t-SNE of the Mouse Dormant HSCs signature. The colour bars indicate log₂-normalized expression. k. The enrichment of GPRC5C^pos-HSCs in clusters 4 and 8 is 20.4% compared to 1.6% for GPRC5C^neg-HSCs. l. t-SNE of stem- and activation-associated genes. The colour bars indicate log₂-normalized expression. m. t-SNE of KEGG and GO pathways. The colour bars indicate log₂-normalized expression. n. Representative FACS histogram of OP-Puro MFI in BM GPRC5C^pos/neg-HSCs. All data presented as mean ± SD. Statistical significance was determined using two-tailed t-test (c, e, f) and at least two independent experiments were performed. ns, not significant. n indicates number of replicates. Source numerical data are available in source data.

Source data