Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Apr 10;14:2018. doi: 10.1038/s41467-023-37783-4

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2023

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/.

PMC Copyright notice

Fig. 8 — a Experimental design for NTN1 treatment in steady state aged mice. b, c Limiting dilution WBM transplantation analysis demonstrating ~4 fold increase in HSC numbers in aged mice treated with NTN1 as compared to PBS treated aged mice, evaluated by number of recipient mice demonstrating long-term multi-lineage reconstitution (LTMR) in their peripheral blood after 16 weeks following transplantation (N = 3 donors/group; N = 20 recipients/group per cell dose). Line graph displaying estimates of HSC frequency in the indicated groups with dashed lines representing 95% confidence intervals. Stem cell frequency and significance were determined using Extreme Limiting Dilution Analysis (ELDA)⁶¹. d, e Competitive HSC transplantation demonstrating that NTN1 treatment of aged mice restores long-term (>4 months) HSC engraftment potential d and myeloid-biased output e, similar to HSCs from young donors (N = 10 donors/group; Recipients N = 20 (Young), N = 44 (Aged-PBS), N = 48 (Aged-NTN1)). f, g Secondary transplantation demonstrating a preservation of serial repopulation f and balanced lineage reconstitution abilities g in donor cells derived from NTN1 treated aged mice (N = 10 donors/group; Recipients N = 10 (Young), N = 20 (Aged-PBS), N = 16 (Aged-NTN1)). h Experimental design describing HSC transplantation strategy to assess the direct effects of NTN1 treatment on aged HSC function (Supplementary Fig. 4d). Following an 11 day ex vivo expansion, 10,000 FACS sorted expansion cells (DAPI^-CD45.2⁺) from each donor were transplanted along with 10⁶ WBM competitor cells (CD45.1) into preconditioned CD45.1 recipients. Following long-term (>6 month) engraftment, CD45.2+ HSCs were FACS purified from primary recipients, and transplanted along with fresh CD45.1 WBM competitor cells into preconditioned CD45.1 secondary recipients. i Bar graphs demonstrating long-term engraftment potential in ex vivo cultured aged HSCs, as compared to HSCs derived from young mice. Note that NTN1 treatment restores LTMR ability of aged HSCs (N = 2 donors/group; Recipients N = 18 (Young), N = 20 (Aged-PBS), N = 19 (Aged-NTN1)). j Bar graphs demonstrating serial repopulation and LTMR (>4 months) (Recipients N = 12 (Young), N = 20 (Aged-NTN1)). Data is presented as the mean ± standard error of the mean (SEM). Statistical significance determined using two-tailed unpaired t-test (for pairwise comparisons), and One-Way ANOVA with Tukey’s correction for multiple comparisons. *P < 0.05; **P < 0.01; ***P < 0.001. ns denotes statistically not significant.