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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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Fig. 6 — a, Experimental design to assess the effect of in vitro HA and NAG treatment on Gprc5c^pos/neg–HSCs. b, SiC division assay of Gprc5c^pos/neg–HSCs quantified after 48 h in vitro culture treatment with NAG, HA and DMSO control. n = 8 biological replicates. c, Transplantation of Gprc5c^pos/neg–HSCs treated with NAG, HA and DMSO control. n = 7 biological replicates, except n = 5 biological replicates for Gprc5c^pos-HSCs treated with NAG. d, Experimental design to assess the effect of in vitro HA and NAG treatment on Gprc5c-KO and WT control HSCs. e, SiC division assay of Gprc5c-KO and WT control HSCs quantified after 48 h in vitro culture treatment with NAG, HA and DMSO control. n = 6 biological replicates. f, Transplantation of Gprc5c-KO and WT control HSCs treated with NAG, HA and DMSO control. n = 9, 9, 6, 11, 12 and 17 biological replicates for WT-DMSO, WT-NAG, WT-HA, Gpr-KO-DMSO, Gpr-KO-NAG and Gpr-KO-HA, respectively. g, Experimental design to assess the effect of in vivo NAG and HA treatment in preventing viral stress from poly I:C. HSCs defined as Lineage⁻cKit⁺Epcr⁺CD150⁺CD48⁻CD34⁻. h, SiC division assay of in vivo treated Gprc5c-KO and WT control HSCs cells quantified after 48 h of in vitro culture. n = 6 biological replicates. i, Differential gene expression. Heat map representing median RNA expression from qPCR data (normalized to housekeeping gene Oaz1 and PBS control) n = 11 biological replicates. j, SiC division kinetics of BM GPRC5C^pos/neg–LT-HSCs with in vitro culture treatment with NAG, HA and DMSO control. n = 4 biological replicates. k, Transplantation of GPRC5C^pos/neg-HSCs from two CB donors treated with HA and control. n = 6 biological replicates. l, Representative summary of findings. All data presented as mean ± s.d., except i where median is presented. Statistical significance was determined using two-tailed t-test (c, f, i and k) or two-way ANOVA (b, e, h and j). 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