Additional file 1 of Thrombopoietin mimetic stimulates bone marrow vascular and stromal niches to mitigate acute radiation syndrome
Creators: name: Vercellino, Justin
name: Małachowska, Beata
name: Kulkarni, Shilpa
name: Bell, Brett I.
name: Shajahan, Shahin
name: Shinoda, Kosaku
name: Eichenbaum, Gary
name: Verma, Amit K.
name: Ghosh, Sanchita P.
name: Yang, Weng-Lang
name: Frenette, Paul S.
name: Guha, Chandan
Publisher: name: figshare
Year: 2024
Subjects: subject: Biophysics
subject: Cell Biology
subject: Molecular Biology
subject: Physiology
subject: FOS: Biological sciences
subject: Pharmacology
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer
subject: Hematology
subject: Computational Biology

description: Additional file 1: Figure S1. TPOm augments the frequency of hematopoietic stem and progenitor, endothelial, and stromal cells in murine bone marrow at homeostasis. (A) Gating strategy and representative flow plots for identifying various hematopoietic stem cell populations for naive and day 3 after TPOm treatment groups. (B) The frequency of live MPC, MkP, LSK, ST-, and LT-HSC of naïve and TPOm-treated mice (n=3-4/group) over time. (C) Gating strategy and representative flow plots for identifying various endothelial cell and stromal populations for naive and day 3 after TPOm treatment groups. (D) The frequency of live EC, EPC, AEC, SEC, and MSC of naïve and TPOm-treated mice (n=3-4/group) over time. Data are expressed as mean ± SEM. *p &lt; 0.05 vs. naïve assessed by one-way ANOVA with post hoc Dunnett test for multiple comparisons. Figure S2. TPOm increases the frequency of hematopoietic stem and progenitor, endothelial, and stromal cells in murine bone marrow following 7.0 Gy sublethal total body irradiation. (A) The frequency of live MPC, MkP, LSK, ST-, and LT-HSC of naïve and TPOm-treated mice (n=4-29/group) over time. (B) The frequency of live EC, EPC, AEC, SEC, and MSC of naïve and TPOm-treated mice (n=4-29/group) over time. Data are expressed as mean ± SEM. *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001 vehicle vs. TPOm-treated assessed by unpaired Student’s t-test with post hoc Holm-Sidak method for multiple comparisons. Outliers were determined using ROUT with a Q = 0.2%. Figure S3. TPOm increases survival of female mice and reduces hemorrhaging in the bone marrow of male mice after lethal total body irradiation.(A) Kaplan-Meier survival curve of vehicle and TPOm treated female mice for 30 days after 7.2 Gy X-ray TBI. (B) The percentage of body weight change over 30 days after 7.2 Gy X-ray TBI in female mice. (C) Representative H&amp;E images of sternal bone marrow from 8.8 Gy (137Cs) irradiated vehicle and TPOm-treated mice on days 7 and 14 after irradiation. For survival the Log-rank (Mantel-Cox) test was used for curve comparison. Light blue arrow, adipocytes; dark blue box, hemorrhaging. Scale bar is 100 μm, For the percent weight change data are expressed as mean ± SEM. Figure S4. TPOm promotes secretion of VEGF-A and VEGF-C in healthy mice. (A, B) ELISA of VEGF-A in (A) serum and in (B) BM after TPOm treatment. (C, D) ELISA of VEGF-C in (C) serum and in (D) BM after TPOm treatment. Data are expressed as mean ± SEM. *p &lt; 0.05 vs. naïve assessed by one-way ANOVA with post hoc Dunnett test for multiple comparisons. Figure S5. TPOm increases Mki67 expression on megakaryocyte progenitors. (A) Cell cycle status of individual UMAP per treatment group. (B) Violin plots of the expression level of Mki67 on megakaryocyte progenitors for each treatment group. Figure S6. MSCs express various collagens and VEGFs. (A) Dot plot of genes highly specific to the MSC cluster of the TNC in the BM. (B) Expression levels of different collagens and their interacting partners on each cluster. (C) Expression levels of different vascular growth factors secreted by each cluster.