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. 2023 Sep 20;14:5850. doi: 10.1038/s41467-023-40923-5

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© 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/.

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Fig. 2 — Panels a and b show representative RBC images from an HU-treated patient collected at 1.7% oxygen tension under shear forces in a microfluidic channel 20 microns wide and 8 microns deep. White arrows indicate the direction the cells were moving when the images were collected. Panel a shows RBCs with no detectable Hb polymer content, while panel b shows RBCs with a detectable polymer content. RBCs from panel a have most of their hemoglobin in the soluble state, and their shape is defined by their position relative to the wall (from left to right: closer to the top wall, at the center, and closer to the bottom wall) of the microfluidic channel and the resulting shear forces. RBCs from panel b have a significant amount of hemoglobin in the polymer state and their morphology (from left to right: elongated, holly leaf, and granular shapes) is altered by the dynamics of polymer aggregation upon deoxygenation. RBCs as shown in panels a and b were collected at 1.7% and 2.7% oxygen tensions for both Untransfused BCL^shmiR and HU High Responders patients but with different fractions. Panels c and d show the fraction of RBCs with no detectable Hb polymer content at 1.7% and 2.7% oxygen tensions, respectively, measured in vitro in Untransfused BCL^shmiR (n = 7; green boxes), in HU High Responders (n = 10; blue boxes), and in HU Low Responders (n = 8; purple boxes). Panels e and f show the HbF percentages (of total Hb) as measured by HPLC, and the proportion of F-cells as measured by flow cytometry. Boxplot properties and the method used to compute the p-values in this study are described in the “Methods” subsection “Data analysis, statistics, and reproducibility”. Source data are provided as a Source Data file.