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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. 8 — a. Serial CFU assay of Gprc5c^pos/neg–HSCs cultured in NAG, HA, or DMSO control for 72 hours. n = 6 biological replicates. b. Left panel: experimental design to assess the effect of in vitro HA, NAG, and DMSO treatment on HA^negGprc5c^pos/neg–HSCs. Right Panel: SiC division assay of HA^negGprc5c^pos/neg–HSCs quantified after 48 hours in vitro culture treatment with NAG, HA, and DMSO. n = 5 biological replicates. c. Serial CFU assay of Gprc5c-KO and WT control HSCs cultured in NAG, HA, and DMSO control for 72 hours. n = 5, 3, 5, 6, 6, and 5 biological replicates for WT-DMSO, WT-NAG, WT-HA, Gpr-KO-DMSO, Gpr-KO-NAG, and Gpr-KO-HA, respectively. d. SiC division assay of GPRC5C^pos/neg–LT-HSCs with in vitro culture treatment for 48 hours with NAG, HA, and DMSO control. n = 4 biological replicates. e. Experimental design to assess the effect of in vitro NAG, HA, and DMSO treatment on CD44-KO and WT control mice. f. Frequency of HSC/MPP populations from 8⁻week-old CD44-KO (n = 6 biological replicates) and WT control (n = 12 biological replicates) mice. g. Differential gene expression. Heatmap representing median RNA expression from qPCR data (normalized to housekeeping gene Oaz1 and WT control). n = 7 biological replicates. h. SiC division assay of CD44-KO and WT control HSCs quantified after 48 hours in vitro culture treatment with NAG, HA and DMSO. n = 15, 15, 13, 8, and 12 biological replicates for WT-DMSO, WT-NAG, WT-HA, CD44-KO-DMSO, CD44-KO-NAG, and CD44-KO-HA, respectively. i. Serial CFU assay of CD44-KO (n = 6 biological replicates) and WT control (n = 10 biological replicates) HSCs cultured in vitro with NAG, HA, and DMSO for 48 hours. All data presented by mean ± SD. Statistical significance was determined using two-tailed t-test (a, c, f, g, i) or two-way ANOVA (b,d, h). ns, not significant. n indicates number of biological replicates. For all experiments, at least two independent experiments were performed. Source numerical data are available in source data.

Source data