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. 2021 Feb 23;12:1248. doi: 10.1038/s41467-021-21451-6

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© The Author(s) 2021

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. 2 — a LSK, LSK Flt3^-, ST-HSCs, and LT-HSCs were examined for the aggresomal marker Thioflavin T (TFT), ER stress markers (GRP78, GRP94, p-PERK, and p-eI2Fα), reactive oxygen species (ROS), and apoptotic markers (annexin V, and cleaved caspase-3) in 12-week-old Slc29a3^+/+ mice pre (day 0) (gray) and post (day 15) (black) irradiation, and Slc29a3^−/− mice pre (day 0) (blue) and post (day 15) (red) irradiation. Data represent mean ± SEM normalized to Slc29a3^+/+ pre (day 0) and post (day 15) irradiation (n = 6 mice/group, except cleaved caspase 3 and annexin V n = 3 mice/group, *p < 0.05 by ANOVA and Tukey’s post hoc test). b The proportion of LT-HSCs in quiescence (G₀) or expressing FOXO3a in 8-week-old Slc29a3^+/+ mice pre (day 0) (gray) and post (day 15) (black) irradiation, and Slc29a3^−/− mice pre (day 0) (blue) and post (day 15) (red) irradiation. Data represent mean ± SEM (n = 6 mice/group, *p < 0.05 by two-tailed t-test). c CHOP/GRP78 ratio in LT-HSCs in 8-week-old Slc29a3^+/+ (black) and Slc29a3^−/− (red) mice pre (day 0) and post (day 15) irradiation. Data represent mean ± SEM (n = 6 mice/group, *p < 0.05 by two-tailed t-test). d Treatment with the PERK inhibitor (GSK2606414, 0.5 mM) (blue) or without (gray) alters cell death in Slc29a3^−/− HSPCs in the presence of tunicamycin (0–2 mg/ml) or thapsigargin (0–2 mM). Data represent mean ± SEM (n = 6 mice/group, *p < 0.05 by two-tailed t-test). e A proportion of LT-HSCs exhibiting high Thioflavin T fluorescence (ThT^high) and ROS positivity (ROS⁺) when cultured in the presence of DMSO (vehicle; red) or salubrinal (10 mM; blue). Data represent mean ± SEM (n = 6, *p < 0.05 by two-tailed t-test). f Proportion of LT-HSCs positive for FOXO3a or cCasp3 when cultured in the presence of DMSO (vehicle; red) or salubrinal (10 mM; blue). Data represent mean ± SEM (n = 6, *p < 0.05 by two-tailed t-test). g Relative number of HSPCs and their subsets positive for GRP78 and ROS in Slc29a3^+/+ and Slc29a3^−/− bone marrow after 6.5 Gy TBI followed by daily treatment vehicle (DMSO; red) or salubrinal (gray; 1 mg/kg, s.c.) and fold change of HSPCs and erythroid cells after 6.5 Gy TBI followed by daily treatment vehicle (DMSO) (red) or salubrinal (gray) (1 mg/kg, s.c.). Data represent mean ± SEM (n = 6, *p < 0.05 by two-tailed t-test). h Survival curves for 60 days were plotted with the Kaplan–Meier method (n = 6/group, ***p < 0.001; Mantel-Cox test). Vehicle (DMSO; red); Sal salubrinal (1 mg/kg, s.c; gray), 4-PBA 4-phenylbutyrate (10 mg/kg, s.c.; blue).