American Society of Hematology

Figure 2.

$FUShigh HSCs exhibit compromised mobility of FUS. (A) Representative FACS plots showing the percentage of FUSlow and FUShigh HSCs (CD34–Flt3–CD150+LSK) in a young (2 months) Fus-gfp mouse. (B) The FACS plot and histogram show the GFP expression in HSCs (CD34–Flt3–CD150+LSK) between young (2 months) and aged (17-18 months) Fus-gfp mice. Data are shown as mean ± SEM, N = 3 mice per group. (C) The FACS plot shows the GFP expression in HSPCs, including CD34–Flt3–LSK (LT-HSC), CD34+Flt3–LSK (ST-HSC), CD34+Flt3+LSK (MPP), CD34+CD16/32– Lineage–Sca-1–c-Kit+ (CMP), CD34+CD16/32+Lineage–Sca-1–c-Kit+ (GMP), CD34–CD16/32– Lineage–Sca-1–c-Kit+ (MEP), and CD127+Flt3+Lineage–Sca-1loc-Kitlo (CLP). (D) The histogram depicts the percentage of FUShigh HSPCs between young (2 months) and aged (17-18 months) Fus-gfp mice. Data are shown as mean ± SEM, N = 3 mice per group. (E) Representative FACS plots showing the percentage of FUSlow and FUShigh HSCs (CD34–LSK) at the end of the third round of transplantation. (F) The histogram depicts the percentage of FUSlow and FUShigh HSCs (CD34–LSK) at the end of the indicated round of transplantation. Data are shown as mean ± SEM, N = 3 to 5 mice per group. (G) Representative FRAP images show the fluorescence recovery of FUS condensates in the cytoplasm of FUSlow and FUShigh LSK cells of 2-month-old Fus-gfp mice upon 0.5 mM ARS treatment for 6 hours. Scale bar represents 2 μm. See also supplemental Video 3. (H) The line plot shows the recovery rate of FUS condensates in FUSlow LSK cells (N = 6) and FUShigh LSK cells (N = 12) of panel G at indicated time points. Data are shown as mean ± SEM. (I) The histograms depict the mobile fraction (99.2% vs 48.9%) and T-half (3.383% vs 63.064%) of FUSlow and FUShigh LSK cells. Data are shown as mean ± SEM. (J) Representative FRAP images show the fluorescence recovery of FUS condensates in the nucleoplasm of FUSlow and FUShigh LSK cells of 2-month-old Fus-gfp mice upon 0.5 mM ARS treatment for 4 hours. Scale bar represents 1 μm. Refer to supplemental Video 4. (K) The line plot shows the recovery rate of FUS condensates in FUSlow LSK cells (N = 19) and FUShigh LSK cells (N = 47) of panel J at indicated time points. Data are shown as mean ± SEM. (L) The histograms depict the mobile fraction (92.2% vs 41.6%) and T-half (4.264% vs 57.8%) of FUSlow and FUShigh LSK cells. Data are shown as mean ± SEM.$

FUS^high HSCs exhibit compromised mobility of FUS. (A) Representative FACS plots showing the percentage of FUS^low and FUS^high HSCs (CD34^–Flt3^–CD150⁺LSK) in a young (2 months) Fus-gfp mouse. (B) The FACS plot and histogram show the GFP expression in HSCs (CD34^–Flt3^–CD150⁺LSK) between young (2 months) and aged (17-18 months) Fus-gfp mice. Data are shown as mean ± SEM, N = 3 mice per group. (C) The FACS plot shows the GFP expression in HSPCs, including CD34^–Flt3^–LSK (LT-HSC), CD34⁺Flt3^–LSK (ST-HSC), CD34⁺Flt3⁺LSK (MPP), CD34⁺CD16/32^– Lineage^–Sca-1^–c-Kit⁺ (CMP), CD34⁺CD16/32⁺Lineage^–Sca-1^–c-Kit⁺ (GMP), CD34^–CD16/32^– Lineage^–Sca-1^–c-Kit⁺ (MEP), and CD127⁺Flt3⁺Lineage^–Sca-1^loc-Kit^lo (CLP). (D) The histogram depicts the percentage of FUS^high HSPCs between young (2 months) and aged (17-18 months) Fus-gfp mice. Data are shown as mean ± SEM, N = 3 mice per group. (E) Representative FACS plots showing the percentage of FUS^low and FUS^high HSCs (CD34^–LSK) at the end of the third round of transplantation. (F) The histogram depicts the percentage of FUS^low and FUS^high HSCs (CD34^–LSK) at the end of the indicated round of transplantation. Data are shown as mean ± SEM, N = 3 to 5 mice per group. (G) Representative FRAP images show the fluorescence recovery of FUS condensates in the cytoplasm of FUS^low and FUS^high LSK cells of 2-month-old Fus-gfp mice upon 0.5 mM ARS treatment for 6 hours. Scale bar represents 2 μm. See also supplemental Video 3. (H) The line plot shows the recovery rate of FUS condensates in FUS^low LSK cells (N = 6) and FUS^high LSK cells (N = 12) of panel G at indicated time points. Data are shown as mean ± SEM. (I) The histograms depict the mobile fraction (99.2% vs 48.9%) and T-half (3.383% vs 63.064%) of FUS^low and FUS^high LSK cells. Data are shown as mean ± SEM. (J) Representative FRAP images show the fluorescence recovery of FUS condensates in the nucleoplasm of FUS^low and FUS^high LSK cells of 2-month-old Fus-gfp mice upon 0.5 mM ARS treatment for 4 hours. Scale bar represents 1 μm. Refer to supplemental Video 4. (K) The line plot shows the recovery rate of FUS condensates in FUS^low LSK cells (N = 19) and FUS^high LSK cells (N = 47) of panel J at indicated time points. Data are shown as mean ± SEM. (L) The histograms depict the mobile fraction (92.2% vs 41.6%) and T-half (4.264% vs 57.8%) of FUS^low and FUS^high LSK cells. Data are shown as mean ± SEM.

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