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hcd45-pe cy7  (Becton Dickinson)

 
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    Becton Dickinson hcd45-pe cy7
    A Schematic presentation of experimental design. One million Molt4-Luc2 cells were transplanted into each sublethally irradiated NSGS recipient. Ten days post bone marrow transplantation (BMT), vehicle, AraC, QC, or AraC + QC were administered daily to the recipients for consecutive 5 days followed by IVIS imaging analysis at the indicated time points. B QC enhances short-term clearance of ALL cells by AraC but failed to prevent ALL relapse in xenografted mice. The distribution (luciferase intensity) of Molt4-Luc2 cells in the mice described in ( A ) were determined by visualizing luciferase using the IVIS system at the indicated time points ( n = 6/group). C QC administration improves the survival of primary recipients transplanted with ALL cells. Survival of the recipients was monitored and plotted by the Kaplan–Meier method (Ctr, n = 8; Vehicle, n = 9; AraC, n = 12; QC, n = 9; AraC+QC, n = 12). Median survival: Vehicle (30 days); AraC (52 days); QC (40 days); and AraC + QC (70 days). D QC enhances the apoptotic effect of AraC on ALL cells in vivo. Human donor-derived cells <t>(hCD45</t> + ) from BM of the recipients described in ( A ) were aspirated and subjected to Flow cytometry analysis for apoptosis at different time points. Representative flow plots (Left) and quantification (Right) are shown. Results are means ± SEM of three independent experiments (AraC, 5 days: n = 10; 15 days: n = 11; AraC + QC, 5 days, n = 11; 15 days, n = 12). See also Supplementary Fig. . Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (** p < 0.01; *** p < 0.001; **** p < 0.0001).
    Hcd45 Pe Cy7, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hcd45-pe cy7/product/Becton Dickinson
    Average 90 stars, based on 1 article reviews
    hcd45-pe cy7 - by Bioz Stars, 2026-03
    90/100 stars

    Images

    1) Product Images from "Quinacrine-CASIN combination overcomes chemoresistance in human acute lymphoid leukemia"

    Article Title: Quinacrine-CASIN combination overcomes chemoresistance in human acute lymphoid leukemia

    Journal: Nature Communications

    doi: 10.1038/s41467-021-27300-w

    A Schematic presentation of experimental design. One million Molt4-Luc2 cells were transplanted into each sublethally irradiated NSGS recipient. Ten days post bone marrow transplantation (BMT), vehicle, AraC, QC, or AraC + QC were administered daily to the recipients for consecutive 5 days followed by IVIS imaging analysis at the indicated time points. B QC enhances short-term clearance of ALL cells by AraC but failed to prevent ALL relapse in xenografted mice. The distribution (luciferase intensity) of Molt4-Luc2 cells in the mice described in ( A ) were determined by visualizing luciferase using the IVIS system at the indicated time points ( n = 6/group). C QC administration improves the survival of primary recipients transplanted with ALL cells. Survival of the recipients was monitored and plotted by the Kaplan–Meier method (Ctr, n = 8; Vehicle, n = 9; AraC, n = 12; QC, n = 9; AraC+QC, n = 12). Median survival: Vehicle (30 days); AraC (52 days); QC (40 days); and AraC + QC (70 days). D QC enhances the apoptotic effect of AraC on ALL cells in vivo. Human donor-derived cells (hCD45 + ) from BM of the recipients described in ( A ) were aspirated and subjected to Flow cytometry analysis for apoptosis at different time points. Representative flow plots (Left) and quantification (Right) are shown. Results are means ± SEM of three independent experiments (AraC, 5 days: n = 10; 15 days: n = 11; AraC + QC, 5 days, n = 11; 15 days, n = 12). See also Supplementary Fig. . Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (** p < 0.01; *** p < 0.001; **** p < 0.0001).
    Figure Legend Snippet: A Schematic presentation of experimental design. One million Molt4-Luc2 cells were transplanted into each sublethally irradiated NSGS recipient. Ten days post bone marrow transplantation (BMT), vehicle, AraC, QC, or AraC + QC were administered daily to the recipients for consecutive 5 days followed by IVIS imaging analysis at the indicated time points. B QC enhances short-term clearance of ALL cells by AraC but failed to prevent ALL relapse in xenografted mice. The distribution (luciferase intensity) of Molt4-Luc2 cells in the mice described in ( A ) were determined by visualizing luciferase using the IVIS system at the indicated time points ( n = 6/group). C QC administration improves the survival of primary recipients transplanted with ALL cells. Survival of the recipients was monitored and plotted by the Kaplan–Meier method (Ctr, n = 8; Vehicle, n = 9; AraC, n = 12; QC, n = 9; AraC+QC, n = 12). Median survival: Vehicle (30 days); AraC (52 days); QC (40 days); and AraC + QC (70 days). D QC enhances the apoptotic effect of AraC on ALL cells in vivo. Human donor-derived cells (hCD45 + ) from BM of the recipients described in ( A ) were aspirated and subjected to Flow cytometry analysis for apoptosis at different time points. Representative flow plots (Left) and quantification (Right) are shown. Results are means ± SEM of three independent experiments (AraC, 5 days: n = 10; 15 days: n = 11; AraC + QC, 5 days, n = 11; 15 days, n = 12). See also Supplementary Fig. . Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Techniques Used: Irradiation, Transplantation Assay, Imaging, Luciferase, In Vivo, Derivative Assay, Flow Cytometry, Two Tailed Test

    A Schematic presentation of experimental design. B QC reduces leukemia burden in primary recipients. Percentage of human cells in the bone marrow of mice transplanted with four ALL patient samples were determined by flow cytometry 1-month post-transplant (ALL2: V, n = 10; n = 8 for all other groups). C , D QC prolongs the survival of primary recipients. Survival of recipients transplanted with ALL2 ( C ) or ALL5 ( D ) cells were monitored and plotted by the Kaplan–Meier method (ALL2: Ctr, n = 8; V, n = 9; AraC; n = 10; QC, n = 8; AraC + QC, n = 9; ALL5: Ctr, n = 8; V, n = 9; AraC, n = 8; QC, n = 12; AraC + QC, n = 12). Median survival of C: Vehicle (35 days); AraC (52 days); QC (42.5 days); and AraC+QC (70 days). Median survival of D: Vehicle (32 days); AraC (55 days); QC (47 days); and AraC+QC (65 days). E , F QC fails to improve survival of secondary transplanted recipients. hCD45 + cells from the primary recipients of the same donor ( C , D ) were sorted and pooled, and transplanted into sublethally irradiated NSGS mice. Survival of recipients transplanted with ALL2 ( E ) or ALL5 ( F ) cells were monitored and plotted by the Kaplan–Meier method (ALL2: Ctr, n = 10; V, n = 12; AraC, n = 12; QC, n = 12; AraC + QC, n = 12; ALL5: Ctr, n = 10; V, n = 12; AraC, n = 13; QC, n = 12; AraC + QC, n = 12). V, Vehicle; A, AraC; QC: Quinacrine; A + QC, AraC + Quinacrine. Median survival of E: Vehicle (25 days); AraC (52 days); QC (37.5 days); and AraC+QC (57.5 days). Median survival of F: Vehicle (25 days); AraC (52 days); QC (35 days); and AraC+QC (55 days). Statistics were performed in the indicated groups: two-sided paired t test (parametric). Animal survival data were analyzed by Gehan–Breslow–Wilcoxon test; p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).
    Figure Legend Snippet: A Schematic presentation of experimental design. B QC reduces leukemia burden in primary recipients. Percentage of human cells in the bone marrow of mice transplanted with four ALL patient samples were determined by flow cytometry 1-month post-transplant (ALL2: V, n = 10; n = 8 for all other groups). C , D QC prolongs the survival of primary recipients. Survival of recipients transplanted with ALL2 ( C ) or ALL5 ( D ) cells were monitored and plotted by the Kaplan–Meier method (ALL2: Ctr, n = 8; V, n = 9; AraC; n = 10; QC, n = 8; AraC + QC, n = 9; ALL5: Ctr, n = 8; V, n = 9; AraC, n = 8; QC, n = 12; AraC + QC, n = 12). Median survival of C: Vehicle (35 days); AraC (52 days); QC (42.5 days); and AraC+QC (70 days). Median survival of D: Vehicle (32 days); AraC (55 days); QC (47 days); and AraC+QC (65 days). E , F QC fails to improve survival of secondary transplanted recipients. hCD45 + cells from the primary recipients of the same donor ( C , D ) were sorted and pooled, and transplanted into sublethally irradiated NSGS mice. Survival of recipients transplanted with ALL2 ( E ) or ALL5 ( F ) cells were monitored and plotted by the Kaplan–Meier method (ALL2: Ctr, n = 10; V, n = 12; AraC, n = 12; QC, n = 12; AraC + QC, n = 12; ALL5: Ctr, n = 10; V, n = 12; AraC, n = 13; QC, n = 12; AraC + QC, n = 12). V, Vehicle; A, AraC; QC: Quinacrine; A + QC, AraC + Quinacrine. Median survival of E: Vehicle (25 days); AraC (52 days); QC (37.5 days); and AraC+QC (57.5 days). Median survival of F: Vehicle (25 days); AraC (52 days); QC (35 days); and AraC+QC (55 days). Statistics were performed in the indicated groups: two-sided paired t test (parametric). Animal survival data were analyzed by Gehan–Breslow–Wilcoxon test; p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Techniques Used: Flow Cytometry, Irradiation

    A Schematic presentation of experimental design. B C dc42 A ctivity S pecific In hibitor, CASIN decreases total hCD45 + cells in the BM of primary recipients. 1–2 × 10 6 primary ALL cells from pooled samples of ALL 1, 4, 7, 8 were transplanted into sublethally irradiated NSGS mice followed by the treatments described in ( A ). BM cells from the recipients were subjected to flow cytometry analysis for hCD45 (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). C CASIN decreases LSC-enriched cell population in the BM of primary recipients. BM cells from recipients described in ( B ) were subjected to flow cytometry analysis for hCD45 and hCD34 (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). D CASIN reduces quiescent leukemia stem cells (LSC)-enriched ALL cells in the BM of the primary recipients. BM cells from recipients described in ( C ) were gated for cell cycle analysis (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). E , F CASIN improves survival of both primary and secondary recipients. BM cells from recipients of the same donor described in ( B ) were pooled and transplanted into sublethally irradiated NSGS recipients. Survival of the primary recipients ( E ; Ctr, n = 8; V, n = 9; AraC, n = 12; AraC + QC, n = 10; AraC + CASIN, n = 9; CASIN + QC + AraC, n = 11) and secondary recipients ( F ; Ctr, n = 10; V, n = 9; AraC, n = 8; AraC + QC, n = 8; AraC + CASIN, n = 8; CASIN + QC + AraC, n = 10) were monitored and plotted by the Kaplan–Meier method. Mice without transplantation served as controls (CTL). V, Vehicle; AraC, AraC; QC: Quinacrine; C, CASIN; AraC+QC, AraC + Quinacrine; AraC+QC + C, CASIN + AraC + Quinacrine. See also Supplementary Fig. . Statistics were performed in the indicated groups: two-sided paired t test (parametric). Animal survival data were analyzed by Gehan–Breslow–Wilcoxon test; p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).
    Figure Legend Snippet: A Schematic presentation of experimental design. B C dc42 A ctivity S pecific In hibitor, CASIN decreases total hCD45 + cells in the BM of primary recipients. 1–2 × 10 6 primary ALL cells from pooled samples of ALL 1, 4, 7, 8 were transplanted into sublethally irradiated NSGS mice followed by the treatments described in ( A ). BM cells from the recipients were subjected to flow cytometry analysis for hCD45 (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). C CASIN decreases LSC-enriched cell population in the BM of primary recipients. BM cells from recipients described in ( B ) were subjected to flow cytometry analysis for hCD45 and hCD34 (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). D CASIN reduces quiescent leukemia stem cells (LSC)-enriched ALL cells in the BM of the primary recipients. BM cells from recipients described in ( C ) were gated for cell cycle analysis (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). E , F CASIN improves survival of both primary and secondary recipients. BM cells from recipients of the same donor described in ( B ) were pooled and transplanted into sublethally irradiated NSGS recipients. Survival of the primary recipients ( E ; Ctr, n = 8; V, n = 9; AraC, n = 12; AraC + QC, n = 10; AraC + CASIN, n = 9; CASIN + QC + AraC, n = 11) and secondary recipients ( F ; Ctr, n = 10; V, n = 9; AraC, n = 8; AraC + QC, n = 8; AraC + CASIN, n = 8; CASIN + QC + AraC, n = 10) were monitored and plotted by the Kaplan–Meier method. Mice without transplantation served as controls (CTL). V, Vehicle; AraC, AraC; QC: Quinacrine; C, CASIN; AraC+QC, AraC + Quinacrine; AraC+QC + C, CASIN + AraC + Quinacrine. See also Supplementary Fig. . Statistics were performed in the indicated groups: two-sided paired t test (parametric). Animal survival data were analyzed by Gehan–Breslow–Wilcoxon test; p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Techniques Used: Irradiation, Flow Cytometry, Cell Cycle Assay, Transplantation Assay

    A Schematic presentation of experimental design. B CASIN does not further increase the cytotoxic effect of AraC on primary ALL cells in vitro. Primary ALL cells were cultured on hTERT-immortalized MSCs in the presence or absence of AraC or AraC + CASIN (AraC, 2 μM; CASIN, 5 μM). Cultured cells were enumerated using counting beads and normalized to the Vehicle controls. Results are means ± SEM of three independent experiments ( n = 6). C Schematic presentation of in vivo experimental design. D CASIN increases donor CD45 + cells in PB and reduces hCD45 + in the bone marrow (BM) of transplanted recipients. Primary ALL cells were transplanted into sublethally irradiated NSGS mice followed by two doses of CASIN injection. Cells from PB (Left) or BM (Right) were subjected to Flow cytometry analysis for hCD45 staining 1-day post treatment. Results are means ± SEM of three independent experiments (PB: V, n = 8; CASIN, n = 6; BM: V, n = 6; CASIN, n = 6). E CASIN decreases quiescent LSC-enriched ALL cells in the BM of transplanted recipients. BM cells described in ( D ) were subjected to Flow cytometry analysis for cell cycle 1 day post treatment. Results are means ± SEM of three independent experiments (PB: V, n = 8; CASIN, n = 6; BM: V, n = 6; CASIN, n = 6). F CASIN increases cycling LSC-enriched ALL cells in the BM of transplanted recipients. BM cells described in ( D ) were subjected to cell cycle analysis 1-day post treatment. Results are means ± SEM of three independent experiments (V, n = 6; CASIN, n = 8). V, Vehicle; AraC, AraC; C, CASIN; AraC + C, AraC + CASIN. Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).
    Figure Legend Snippet: A Schematic presentation of experimental design. B CASIN does not further increase the cytotoxic effect of AraC on primary ALL cells in vitro. Primary ALL cells were cultured on hTERT-immortalized MSCs in the presence or absence of AraC or AraC + CASIN (AraC, 2 μM; CASIN, 5 μM). Cultured cells were enumerated using counting beads and normalized to the Vehicle controls. Results are means ± SEM of three independent experiments ( n = 6). C Schematic presentation of in vivo experimental design. D CASIN increases donor CD45 + cells in PB and reduces hCD45 + in the bone marrow (BM) of transplanted recipients. Primary ALL cells were transplanted into sublethally irradiated NSGS mice followed by two doses of CASIN injection. Cells from PB (Left) or BM (Right) were subjected to Flow cytometry analysis for hCD45 staining 1-day post treatment. Results are means ± SEM of three independent experiments (PB: V, n = 8; CASIN, n = 6; BM: V, n = 6; CASIN, n = 6). E CASIN decreases quiescent LSC-enriched ALL cells in the BM of transplanted recipients. BM cells described in ( D ) were subjected to Flow cytometry analysis for cell cycle 1 day post treatment. Results are means ± SEM of three independent experiments (PB: V, n = 8; CASIN, n = 6; BM: V, n = 6; CASIN, n = 6). F CASIN increases cycling LSC-enriched ALL cells in the BM of transplanted recipients. BM cells described in ( D ) were subjected to cell cycle analysis 1-day post treatment. Results are means ± SEM of three independent experiments (V, n = 6; CASIN, n = 8). V, Vehicle; AraC, AraC; C, CASIN; AraC + C, AraC + CASIN. Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Techniques Used: In Vitro, Cell Culture, In Vivo, Irradiation, Injection, Flow Cytometry, Staining, Cell Cycle Assay, Two Tailed Test

    A Schematic presentation of experimental design. B The effect of CASIN + QC + AraC treatment on total human engraftment in transplanted recipients. 1–3 × 10 4 hCB CD34 + cells were transplanted into sublethally irradiated NSGS mice followed by the treatment described in ( A ). Percentages of human cells (hCD45 + ) in the transplanted recipients 1 week and 8 weeks post treatment were determined by flow cytometry. Results are means ± SEM of three independent experiments (Week 1: V, n = 10; AraC, n = 8; QC, n = 10; AraC + QC, n = 8; CASIN + QC + AraC, n = 8; Week 8: V, n = 8; AraC, n = 9; QC, n = 9; AraC + QC, n = 8; CASIN + QC + AraC, n = 8). C , D The effect of CASIN + QC + AraC treatment on human HSPCs. BM cells were aspirated from the transplanted recipients described in ( B ), and subjected to Flow cytometry analysis for HSPCs (hCD45 + CD34 + CD38 − cells; C ) and HPCs (hCD45 + CD34 + CD38 + cells; D ) 1 week and 8 weeks post treatment. Results are means ± SEM of three independent experiments (Week 1: V, n = 10; AraC, n = 8; QC, n = 8; AraC + QC, n = 8; CASIN + QC + AraC, n = 8; Week 8: V, n = 8; AraC, n = 8; QC, n = 9; AraC + QC, n = 8; CASIN + QC + AraC, n = 9). E CASIN + QC + AraC treatment does not affect long-term repopulating capacity of human HSPCs. BM cells from the primary recipients described in ( B ) were pooled for secondary transplantation to sublethally irradiated NSGS mice. Human engraftment was determined by Flow cytometry. Results are means ± SEM of three independent experiments ( n = 8 per group). V, Vehicle; AraC, AraC; QC: Quinacrine; C, CASIN; AraC + QC, AraC + Quinacrine; AraC + QC + C, CASIN + AraC + Quinacrine. Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (* p < 0.05; ** p < 0.01).
    Figure Legend Snippet: A Schematic presentation of experimental design. B The effect of CASIN + QC + AraC treatment on total human engraftment in transplanted recipients. 1–3 × 10 4 hCB CD34 + cells were transplanted into sublethally irradiated NSGS mice followed by the treatment described in ( A ). Percentages of human cells (hCD45 + ) in the transplanted recipients 1 week and 8 weeks post treatment were determined by flow cytometry. Results are means ± SEM of three independent experiments (Week 1: V, n = 10; AraC, n = 8; QC, n = 10; AraC + QC, n = 8; CASIN + QC + AraC, n = 8; Week 8: V, n = 8; AraC, n = 9; QC, n = 9; AraC + QC, n = 8; CASIN + QC + AraC, n = 8). C , D The effect of CASIN + QC + AraC treatment on human HSPCs. BM cells were aspirated from the transplanted recipients described in ( B ), and subjected to Flow cytometry analysis for HSPCs (hCD45 + CD34 + CD38 − cells; C ) and HPCs (hCD45 + CD34 + CD38 + cells; D ) 1 week and 8 weeks post treatment. Results are means ± SEM of three independent experiments (Week 1: V, n = 10; AraC, n = 8; QC, n = 8; AraC + QC, n = 8; CASIN + QC + AraC, n = 8; Week 8: V, n = 8; AraC, n = 8; QC, n = 9; AraC + QC, n = 8; CASIN + QC + AraC, n = 9). E CASIN + QC + AraC treatment does not affect long-term repopulating capacity of human HSPCs. BM cells from the primary recipients described in ( B ) were pooled for secondary transplantation to sublethally irradiated NSGS mice. Human engraftment was determined by Flow cytometry. Results are means ± SEM of three independent experiments ( n = 8 per group). V, Vehicle; AraC, AraC; QC: Quinacrine; C, CASIN; AraC + QC, AraC + Quinacrine; AraC + QC + C, CASIN + AraC + Quinacrine. Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (* p < 0.05; ** p < 0.01).

    Techniques Used: Irradiation, Flow Cytometry, Transplantation Assay, Two Tailed Test



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    A Schematic presentation of experimental design. One million Molt4-Luc2 cells were transplanted into each sublethally irradiated NSGS recipient. Ten days post bone marrow transplantation (BMT), vehicle, AraC, QC, or AraC + QC were administered daily to the recipients for consecutive 5 days followed by IVIS imaging analysis at the indicated time points. B QC enhances short-term clearance of ALL cells by AraC but failed to prevent ALL relapse in xenografted mice. The distribution (luciferase intensity) of Molt4-Luc2 cells in the mice described in ( A ) were determined by visualizing luciferase using the IVIS system at the indicated time points ( n = 6/group). C QC administration improves the survival of primary recipients transplanted with ALL cells. Survival of the recipients was monitored and plotted by the Kaplan–Meier method (Ctr, n = 8; Vehicle, n = 9; AraC, n = 12; QC, n = 9; AraC+QC, n = 12). Median survival: Vehicle (30 days); AraC (52 days); QC (40 days); and AraC + QC (70 days). D QC enhances the apoptotic effect of AraC on ALL cells in vivo. Human donor-derived cells <t>(hCD45</t> + ) from BM of the recipients described in ( A ) were aspirated and subjected to Flow cytometry analysis for apoptosis at different time points. Representative flow plots (Left) and quantification (Right) are shown. Results are means ± SEM of three independent experiments (AraC, 5 days: n = 10; 15 days: n = 11; AraC + QC, 5 days, n = 11; 15 days, n = 12). See also Supplementary Fig. . Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (** p < 0.01; *** p < 0.001; **** p < 0.0001).
    Hcd45 Pe Cy7, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    (A) Experimental scheme for GGF and 3GF LDA using AFT024 monolayers. (B) ELDA of GGF and 3GF cells after 5 weeks of AFT024 LTC, with a 95% CI of 1/2740 – 1/5899 (frequency of 1/4020) for 3GF and 1/4834 – 1/11527 (frequency of 1/7465) for GGF. (C) Experimental scheme for intrahepatic transplants of D15 CD49f+ sorted 3GF cells into newborn NSG mice. (D) Representative flow cytometry contour plots of <t>hCD45</t> chimerism in CB controls and 2 mice that showed engraftment using D15 3GF cells. (E) Representative flow cytometry contour plots showing the multilineage distribution of week 8 hCD45+ cells from CB controls and D15 3GF IH transplants. CD3+ T cells, CD19+ B cells and CD11c+ myeloid cells. PBMC = peripheral blood mononclear cells and were used as a staining control (non-transplanted).
    Pe Cy7 Hcd45, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti-hcd45-pe-cy7  (Thermo Fisher)
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    (A) Experimental scheme for GGF and 3GF LDA using AFT024 monolayers. (B) ELDA of GGF and 3GF cells after 5 weeks of AFT024 LTC, with a 95% CI of 1/2740 – 1/5899 (frequency of 1/4020) for 3GF and 1/4834 – 1/11527 (frequency of 1/7465) for GGF. (C) Experimental scheme for intrahepatic transplants of D15 CD49f+ sorted 3GF cells into newborn NSG mice. (D) Representative flow cytometry contour plots of <t>hCD45</t> chimerism in CB controls and 2 mice that showed engraftment using D15 3GF cells. (E) Representative flow cytometry contour plots showing the multilineage distribution of week 8 hCD45+ cells from CB controls and D15 3GF IH transplants. CD3+ T cells, CD19+ B cells and CD11c+ myeloid cells. PBMC = peripheral blood mononclear cells and were used as a staining control (non-transplanted).
    Anti Hcd45 Pe Cy7, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Diagram depicting timelines for in vivo experiments. b. Engraftment in blood, bone marrow and spleen in NSG cohorts, measured as the proportion of human cells (hCD45+). c. Engraftment in blood, bone marrow and spleen in SGM3 cohorts, measured as the proportion of human cells (hCD45+). d. Percentage of myeloid, B cell lymphoid, T cell lymphoid and HSPC in the bone marrow of NSG cohorts. e. Percentage of myeloid, B cell lymphoid, T cell lymphoid and HSPC in the bone marrow of SGM3 cohorts. N/A: Not analyzed. Empty circles = SGM3. Filled circles = NSG. Data are represented as mean ± standard deviation. Statistical comparisons were performed using one way ANOVA followed by a Tukey’s Honestly-Significant Difference post-hoc test between each two groups; Tukey’s p values are only shown if ANOVA was significant; ns= non-significant, *p≤0.05, **p≤0.01.

    Journal: bioRxiv

    Article Title: TALEN-mediated intron editing of HSPCs enables transgene expression restricted to the myeloid lineage

    doi: 10.1101/2024.03.05.583596

    Figure Lengend Snippet: Diagram depicting timelines for in vivo experiments. b. Engraftment in blood, bone marrow and spleen in NSG cohorts, measured as the proportion of human cells (hCD45+). c. Engraftment in blood, bone marrow and spleen in SGM3 cohorts, measured as the proportion of human cells (hCD45+). d. Percentage of myeloid, B cell lymphoid, T cell lymphoid and HSPC in the bone marrow of NSG cohorts. e. Percentage of myeloid, B cell lymphoid, T cell lymphoid and HSPC in the bone marrow of SGM3 cohorts. N/A: Not analyzed. Empty circles = SGM3. Filled circles = NSG. Data are represented as mean ± standard deviation. Statistical comparisons were performed using one way ANOVA followed by a Tukey’s Honestly-Significant Difference post-hoc test between each two groups; Tukey’s p values are only shown if ANOVA was significant; ns= non-significant, *p≤0.05, **p≤0.01.

    Article Snippet: For neurons and neuroglia from both eGFP-edited and IDUA- edited mouse cohorts, the following antibody panel was used: hCD45-PE-Cy7 (Clone: REA747; Miltenyi), mCD45-APC-Cy7 (Clone: 30-F11; BD Biosciences), and CD11b-PE (Clone: M1/70; Biolegend).

    Techniques: In Vivo, Standard Deviation

    Edited HSPCs as a vehicle for therapeutic delivery across the blood-brain barrier. a. Engraftment in brain, measured as the proportion of human cells (hCD45+) in the leukocyte compartment in NSG and SGM3 animals injected with HSPC edited with the eGFP or IDUA cassettes. b. Percentage of CD11b + among the human cells from animals injected with HSPC edited with the eGFP or IDUA cassettes. c. Percentage of eGFP+ cells within the human fraction of cell isolated from the brain of animals engrafted with HSPC edited with eGFP cassettes. d. Percentage of eGFP+ cells within the human CD11b + and CD11b - fractions from the brain of animals engrafted with HSPC edited with eGFP cassettes. e. Percentage of edited alleles among human cells found in the brain of animals engrafted with HSPC edited with IDUA cassettes. Empty circles = SGM3. Filled circles = NSG. Data are represented as mean ± standard deviation. Statistical comparisons were performed using unpaired T tests when comparing two groups , or one way ANOVA when comparing three groups followed by a Tukey’s Honestly- Significant Difference post-hoc test between each two groups ( , 5b). Tukey’s p values are only shown if ANOVA was significant; ns= non-significant, *p≤0.05.

    Journal: bioRxiv

    Article Title: TALEN-mediated intron editing of HSPCs enables transgene expression restricted to the myeloid lineage

    doi: 10.1101/2024.03.05.583596

    Figure Lengend Snippet: Edited HSPCs as a vehicle for therapeutic delivery across the blood-brain barrier. a. Engraftment in brain, measured as the proportion of human cells (hCD45+) in the leukocyte compartment in NSG and SGM3 animals injected with HSPC edited with the eGFP or IDUA cassettes. b. Percentage of CD11b + among the human cells from animals injected with HSPC edited with the eGFP or IDUA cassettes. c. Percentage of eGFP+ cells within the human fraction of cell isolated from the brain of animals engrafted with HSPC edited with eGFP cassettes. d. Percentage of eGFP+ cells within the human CD11b + and CD11b - fractions from the brain of animals engrafted with HSPC edited with eGFP cassettes. e. Percentage of edited alleles among human cells found in the brain of animals engrafted with HSPC edited with IDUA cassettes. Empty circles = SGM3. Filled circles = NSG. Data are represented as mean ± standard deviation. Statistical comparisons were performed using unpaired T tests when comparing two groups , or one way ANOVA when comparing three groups followed by a Tukey’s Honestly- Significant Difference post-hoc test between each two groups ( , 5b). Tukey’s p values are only shown if ANOVA was significant; ns= non-significant, *p≤0.05.

    Article Snippet: For neurons and neuroglia from both eGFP-edited and IDUA- edited mouse cohorts, the following antibody panel was used: hCD45-PE-Cy7 (Clone: REA747; Miltenyi), mCD45-APC-Cy7 (Clone: 30-F11; BD Biosciences), and CD11b-PE (Clone: M1/70; Biolegend).

    Techniques: Injection, Isolation, Standard Deviation

    A Schematic presentation of experimental design. One million Molt4-Luc2 cells were transplanted into each sublethally irradiated NSGS recipient. Ten days post bone marrow transplantation (BMT), vehicle, AraC, QC, or AraC + QC were administered daily to the recipients for consecutive 5 days followed by IVIS imaging analysis at the indicated time points. B QC enhances short-term clearance of ALL cells by AraC but failed to prevent ALL relapse in xenografted mice. The distribution (luciferase intensity) of Molt4-Luc2 cells in the mice described in ( A ) were determined by visualizing luciferase using the IVIS system at the indicated time points ( n = 6/group). C QC administration improves the survival of primary recipients transplanted with ALL cells. Survival of the recipients was monitored and plotted by the Kaplan–Meier method (Ctr, n = 8; Vehicle, n = 9; AraC, n = 12; QC, n = 9; AraC+QC, n = 12). Median survival: Vehicle (30 days); AraC (52 days); QC (40 days); and AraC + QC (70 days). D QC enhances the apoptotic effect of AraC on ALL cells in vivo. Human donor-derived cells (hCD45 + ) from BM of the recipients described in ( A ) were aspirated and subjected to Flow cytometry analysis for apoptosis at different time points. Representative flow plots (Left) and quantification (Right) are shown. Results are means ± SEM of three independent experiments (AraC, 5 days: n = 10; 15 days: n = 11; AraC + QC, 5 days, n = 11; 15 days, n = 12). See also Supplementary Fig. . Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Journal: Nature Communications

    Article Title: Quinacrine-CASIN combination overcomes chemoresistance in human acute lymphoid leukemia

    doi: 10.1038/s41467-021-27300-w

    Figure Lengend Snippet: A Schematic presentation of experimental design. One million Molt4-Luc2 cells were transplanted into each sublethally irradiated NSGS recipient. Ten days post bone marrow transplantation (BMT), vehicle, AraC, QC, or AraC + QC were administered daily to the recipients for consecutive 5 days followed by IVIS imaging analysis at the indicated time points. B QC enhances short-term clearance of ALL cells by AraC but failed to prevent ALL relapse in xenografted mice. The distribution (luciferase intensity) of Molt4-Luc2 cells in the mice described in ( A ) were determined by visualizing luciferase using the IVIS system at the indicated time points ( n = 6/group). C QC administration improves the survival of primary recipients transplanted with ALL cells. Survival of the recipients was monitored and plotted by the Kaplan–Meier method (Ctr, n = 8; Vehicle, n = 9; AraC, n = 12; QC, n = 9; AraC+QC, n = 12). Median survival: Vehicle (30 days); AraC (52 days); QC (40 days); and AraC + QC (70 days). D QC enhances the apoptotic effect of AraC on ALL cells in vivo. Human donor-derived cells (hCD45 + ) from BM of the recipients described in ( A ) were aspirated and subjected to Flow cytometry analysis for apoptosis at different time points. Representative flow plots (Left) and quantification (Right) are shown. Results are means ± SEM of three independent experiments (AraC, 5 days: n = 10; 15 days: n = 11; AraC + QC, 5 days, n = 11; 15 days, n = 12). See also Supplementary Fig. . Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Article Snippet: For analysis and sorting of ALL and HSPCs derived from human Cord Blood or adult BM, cells were stained with hCD45-PE Cy7 (Clone: H30, Cat: 560915; BD Biosciences, San Jose, CA), mCD45-PerCP Cy5.5 (Clone:30-F11, Cat: 550994; BD Biosciences, San Jose, CA), Lineage-FITC (Lin1, Cat: 340546; BD Biosciences, San Jose, CA), CD34-PE (Clone 581, Cat: 560941, dilution: 1 in 25; BD Biosciences, San Jose, CA), and CD38-APC (Clone HIT2, Cat: 555462; BD Biosciences, San Jose, CA).

    Techniques: Irradiation, Transplantation Assay, Imaging, Luciferase, In Vivo, Derivative Assay, Flow Cytometry, Two Tailed Test

    A Schematic presentation of experimental design. B QC reduces leukemia burden in primary recipients. Percentage of human cells in the bone marrow of mice transplanted with four ALL patient samples were determined by flow cytometry 1-month post-transplant (ALL2: V, n = 10; n = 8 for all other groups). C , D QC prolongs the survival of primary recipients. Survival of recipients transplanted with ALL2 ( C ) or ALL5 ( D ) cells were monitored and plotted by the Kaplan–Meier method (ALL2: Ctr, n = 8; V, n = 9; AraC; n = 10; QC, n = 8; AraC + QC, n = 9; ALL5: Ctr, n = 8; V, n = 9; AraC, n = 8; QC, n = 12; AraC + QC, n = 12). Median survival of C: Vehicle (35 days); AraC (52 days); QC (42.5 days); and AraC+QC (70 days). Median survival of D: Vehicle (32 days); AraC (55 days); QC (47 days); and AraC+QC (65 days). E , F QC fails to improve survival of secondary transplanted recipients. hCD45 + cells from the primary recipients of the same donor ( C , D ) were sorted and pooled, and transplanted into sublethally irradiated NSGS mice. Survival of recipients transplanted with ALL2 ( E ) or ALL5 ( F ) cells were monitored and plotted by the Kaplan–Meier method (ALL2: Ctr, n = 10; V, n = 12; AraC, n = 12; QC, n = 12; AraC + QC, n = 12; ALL5: Ctr, n = 10; V, n = 12; AraC, n = 13; QC, n = 12; AraC + QC, n = 12). V, Vehicle; A, AraC; QC: Quinacrine; A + QC, AraC + Quinacrine. Median survival of E: Vehicle (25 days); AraC (52 days); QC (37.5 days); and AraC+QC (57.5 days). Median survival of F: Vehicle (25 days); AraC (52 days); QC (35 days); and AraC+QC (55 days). Statistics were performed in the indicated groups: two-sided paired t test (parametric). Animal survival data were analyzed by Gehan–Breslow–Wilcoxon test; p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Journal: Nature Communications

    Article Title: Quinacrine-CASIN combination overcomes chemoresistance in human acute lymphoid leukemia

    doi: 10.1038/s41467-021-27300-w

    Figure Lengend Snippet: A Schematic presentation of experimental design. B QC reduces leukemia burden in primary recipients. Percentage of human cells in the bone marrow of mice transplanted with four ALL patient samples were determined by flow cytometry 1-month post-transplant (ALL2: V, n = 10; n = 8 for all other groups). C , D QC prolongs the survival of primary recipients. Survival of recipients transplanted with ALL2 ( C ) or ALL5 ( D ) cells were monitored and plotted by the Kaplan–Meier method (ALL2: Ctr, n = 8; V, n = 9; AraC; n = 10; QC, n = 8; AraC + QC, n = 9; ALL5: Ctr, n = 8; V, n = 9; AraC, n = 8; QC, n = 12; AraC + QC, n = 12). Median survival of C: Vehicle (35 days); AraC (52 days); QC (42.5 days); and AraC+QC (70 days). Median survival of D: Vehicle (32 days); AraC (55 days); QC (47 days); and AraC+QC (65 days). E , F QC fails to improve survival of secondary transplanted recipients. hCD45 + cells from the primary recipients of the same donor ( C , D ) were sorted and pooled, and transplanted into sublethally irradiated NSGS mice. Survival of recipients transplanted with ALL2 ( E ) or ALL5 ( F ) cells were monitored and plotted by the Kaplan–Meier method (ALL2: Ctr, n = 10; V, n = 12; AraC, n = 12; QC, n = 12; AraC + QC, n = 12; ALL5: Ctr, n = 10; V, n = 12; AraC, n = 13; QC, n = 12; AraC + QC, n = 12). V, Vehicle; A, AraC; QC: Quinacrine; A + QC, AraC + Quinacrine. Median survival of E: Vehicle (25 days); AraC (52 days); QC (37.5 days); and AraC+QC (57.5 days). Median survival of F: Vehicle (25 days); AraC (52 days); QC (35 days); and AraC+QC (55 days). Statistics were performed in the indicated groups: two-sided paired t test (parametric). Animal survival data were analyzed by Gehan–Breslow–Wilcoxon test; p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Article Snippet: For analysis and sorting of ALL and HSPCs derived from human Cord Blood or adult BM, cells were stained with hCD45-PE Cy7 (Clone: H30, Cat: 560915; BD Biosciences, San Jose, CA), mCD45-PerCP Cy5.5 (Clone:30-F11, Cat: 550994; BD Biosciences, San Jose, CA), Lineage-FITC (Lin1, Cat: 340546; BD Biosciences, San Jose, CA), CD34-PE (Clone 581, Cat: 560941, dilution: 1 in 25; BD Biosciences, San Jose, CA), and CD38-APC (Clone HIT2, Cat: 555462; BD Biosciences, San Jose, CA).

    Techniques: Flow Cytometry, Irradiation

    A Schematic presentation of experimental design. B C dc42 A ctivity S pecific In hibitor, CASIN decreases total hCD45 + cells in the BM of primary recipients. 1–2 × 10 6 primary ALL cells from pooled samples of ALL 1, 4, 7, 8 were transplanted into sublethally irradiated NSGS mice followed by the treatments described in ( A ). BM cells from the recipients were subjected to flow cytometry analysis for hCD45 (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). C CASIN decreases LSC-enriched cell population in the BM of primary recipients. BM cells from recipients described in ( B ) were subjected to flow cytometry analysis for hCD45 and hCD34 (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). D CASIN reduces quiescent leukemia stem cells (LSC)-enriched ALL cells in the BM of the primary recipients. BM cells from recipients described in ( C ) were gated for cell cycle analysis (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). E , F CASIN improves survival of both primary and secondary recipients. BM cells from recipients of the same donor described in ( B ) were pooled and transplanted into sublethally irradiated NSGS recipients. Survival of the primary recipients ( E ; Ctr, n = 8; V, n = 9; AraC, n = 12; AraC + QC, n = 10; AraC + CASIN, n = 9; CASIN + QC + AraC, n = 11) and secondary recipients ( F ; Ctr, n = 10; V, n = 9; AraC, n = 8; AraC + QC, n = 8; AraC + CASIN, n = 8; CASIN + QC + AraC, n = 10) were monitored and plotted by the Kaplan–Meier method. Mice without transplantation served as controls (CTL). V, Vehicle; AraC, AraC; QC: Quinacrine; C, CASIN; AraC+QC, AraC + Quinacrine; AraC+QC + C, CASIN + AraC + Quinacrine. See also Supplementary Fig. . Statistics were performed in the indicated groups: two-sided paired t test (parametric). Animal survival data were analyzed by Gehan–Breslow–Wilcoxon test; p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Journal: Nature Communications

    Article Title: Quinacrine-CASIN combination overcomes chemoresistance in human acute lymphoid leukemia

    doi: 10.1038/s41467-021-27300-w

    Figure Lengend Snippet: A Schematic presentation of experimental design. B C dc42 A ctivity S pecific In hibitor, CASIN decreases total hCD45 + cells in the BM of primary recipients. 1–2 × 10 6 primary ALL cells from pooled samples of ALL 1, 4, 7, 8 were transplanted into sublethally irradiated NSGS mice followed by the treatments described in ( A ). BM cells from the recipients were subjected to flow cytometry analysis for hCD45 (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). C CASIN decreases LSC-enriched cell population in the BM of primary recipients. BM cells from recipients described in ( B ) were subjected to flow cytometry analysis for hCD45 and hCD34 (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). D CASIN reduces quiescent leukemia stem cells (LSC)-enriched ALL cells in the BM of the primary recipients. BM cells from recipients described in ( C ) were gated for cell cycle analysis (V, n = 10; AraC, n = 9; AraC + QC, n = 8, CASIN + QC + AraC, n = 8). E , F CASIN improves survival of both primary and secondary recipients. BM cells from recipients of the same donor described in ( B ) were pooled and transplanted into sublethally irradiated NSGS recipients. Survival of the primary recipients ( E ; Ctr, n = 8; V, n = 9; AraC, n = 12; AraC + QC, n = 10; AraC + CASIN, n = 9; CASIN + QC + AraC, n = 11) and secondary recipients ( F ; Ctr, n = 10; V, n = 9; AraC, n = 8; AraC + QC, n = 8; AraC + CASIN, n = 8; CASIN + QC + AraC, n = 10) were monitored and plotted by the Kaplan–Meier method. Mice without transplantation served as controls (CTL). V, Vehicle; AraC, AraC; QC: Quinacrine; C, CASIN; AraC+QC, AraC + Quinacrine; AraC+QC + C, CASIN + AraC + Quinacrine. See also Supplementary Fig. . Statistics were performed in the indicated groups: two-sided paired t test (parametric). Animal survival data were analyzed by Gehan–Breslow–Wilcoxon test; p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Article Snippet: For analysis and sorting of ALL and HSPCs derived from human Cord Blood or adult BM, cells were stained with hCD45-PE Cy7 (Clone: H30, Cat: 560915; BD Biosciences, San Jose, CA), mCD45-PerCP Cy5.5 (Clone:30-F11, Cat: 550994; BD Biosciences, San Jose, CA), Lineage-FITC (Lin1, Cat: 340546; BD Biosciences, San Jose, CA), CD34-PE (Clone 581, Cat: 560941, dilution: 1 in 25; BD Biosciences, San Jose, CA), and CD38-APC (Clone HIT2, Cat: 555462; BD Biosciences, San Jose, CA).

    Techniques: Irradiation, Flow Cytometry, Cell Cycle Assay, Transplantation Assay

    A Schematic presentation of experimental design. B CASIN does not further increase the cytotoxic effect of AraC on primary ALL cells in vitro. Primary ALL cells were cultured on hTERT-immortalized MSCs in the presence or absence of AraC or AraC + CASIN (AraC, 2 μM; CASIN, 5 μM). Cultured cells were enumerated using counting beads and normalized to the Vehicle controls. Results are means ± SEM of three independent experiments ( n = 6). C Schematic presentation of in vivo experimental design. D CASIN increases donor CD45 + cells in PB and reduces hCD45 + in the bone marrow (BM) of transplanted recipients. Primary ALL cells were transplanted into sublethally irradiated NSGS mice followed by two doses of CASIN injection. Cells from PB (Left) or BM (Right) were subjected to Flow cytometry analysis for hCD45 staining 1-day post treatment. Results are means ± SEM of three independent experiments (PB: V, n = 8; CASIN, n = 6; BM: V, n = 6; CASIN, n = 6). E CASIN decreases quiescent LSC-enriched ALL cells in the BM of transplanted recipients. BM cells described in ( D ) were subjected to Flow cytometry analysis for cell cycle 1 day post treatment. Results are means ± SEM of three independent experiments (PB: V, n = 8; CASIN, n = 6; BM: V, n = 6; CASIN, n = 6). F CASIN increases cycling LSC-enriched ALL cells in the BM of transplanted recipients. BM cells described in ( D ) were subjected to cell cycle analysis 1-day post treatment. Results are means ± SEM of three independent experiments (V, n = 6; CASIN, n = 8). V, Vehicle; AraC, AraC; C, CASIN; AraC + C, AraC + CASIN. Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Journal: Nature Communications

    Article Title: Quinacrine-CASIN combination overcomes chemoresistance in human acute lymphoid leukemia

    doi: 10.1038/s41467-021-27300-w

    Figure Lengend Snippet: A Schematic presentation of experimental design. B CASIN does not further increase the cytotoxic effect of AraC on primary ALL cells in vitro. Primary ALL cells were cultured on hTERT-immortalized MSCs in the presence or absence of AraC or AraC + CASIN (AraC, 2 μM; CASIN, 5 μM). Cultured cells were enumerated using counting beads and normalized to the Vehicle controls. Results are means ± SEM of three independent experiments ( n = 6). C Schematic presentation of in vivo experimental design. D CASIN increases donor CD45 + cells in PB and reduces hCD45 + in the bone marrow (BM) of transplanted recipients. Primary ALL cells were transplanted into sublethally irradiated NSGS mice followed by two doses of CASIN injection. Cells from PB (Left) or BM (Right) were subjected to Flow cytometry analysis for hCD45 staining 1-day post treatment. Results are means ± SEM of three independent experiments (PB: V, n = 8; CASIN, n = 6; BM: V, n = 6; CASIN, n = 6). E CASIN decreases quiescent LSC-enriched ALL cells in the BM of transplanted recipients. BM cells described in ( D ) were subjected to Flow cytometry analysis for cell cycle 1 day post treatment. Results are means ± SEM of three independent experiments (PB: V, n = 8; CASIN, n = 6; BM: V, n = 6; CASIN, n = 6). F CASIN increases cycling LSC-enriched ALL cells in the BM of transplanted recipients. BM cells described in ( D ) were subjected to cell cycle analysis 1-day post treatment. Results are means ± SEM of three independent experiments (V, n = 6; CASIN, n = 8). V, Vehicle; AraC, AraC; C, CASIN; AraC + C, AraC + CASIN. Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

    Article Snippet: For analysis and sorting of ALL and HSPCs derived from human Cord Blood or adult BM, cells were stained with hCD45-PE Cy7 (Clone: H30, Cat: 560915; BD Biosciences, San Jose, CA), mCD45-PerCP Cy5.5 (Clone:30-F11, Cat: 550994; BD Biosciences, San Jose, CA), Lineage-FITC (Lin1, Cat: 340546; BD Biosciences, San Jose, CA), CD34-PE (Clone 581, Cat: 560941, dilution: 1 in 25; BD Biosciences, San Jose, CA), and CD38-APC (Clone HIT2, Cat: 555462; BD Biosciences, San Jose, CA).

    Techniques: In Vitro, Cell Culture, In Vivo, Irradiation, Injection, Flow Cytometry, Staining, Cell Cycle Assay, Two Tailed Test

    A Schematic presentation of experimental design. B The effect of CASIN + QC + AraC treatment on total human engraftment in transplanted recipients. 1–3 × 10 4 hCB CD34 + cells were transplanted into sublethally irradiated NSGS mice followed by the treatment described in ( A ). Percentages of human cells (hCD45 + ) in the transplanted recipients 1 week and 8 weeks post treatment were determined by flow cytometry. Results are means ± SEM of three independent experiments (Week 1: V, n = 10; AraC, n = 8; QC, n = 10; AraC + QC, n = 8; CASIN + QC + AraC, n = 8; Week 8: V, n = 8; AraC, n = 9; QC, n = 9; AraC + QC, n = 8; CASIN + QC + AraC, n = 8). C , D The effect of CASIN + QC + AraC treatment on human HSPCs. BM cells were aspirated from the transplanted recipients described in ( B ), and subjected to Flow cytometry analysis for HSPCs (hCD45 + CD34 + CD38 − cells; C ) and HPCs (hCD45 + CD34 + CD38 + cells; D ) 1 week and 8 weeks post treatment. Results are means ± SEM of three independent experiments (Week 1: V, n = 10; AraC, n = 8; QC, n = 8; AraC + QC, n = 8; CASIN + QC + AraC, n = 8; Week 8: V, n = 8; AraC, n = 8; QC, n = 9; AraC + QC, n = 8; CASIN + QC + AraC, n = 9). E CASIN + QC + AraC treatment does not affect long-term repopulating capacity of human HSPCs. BM cells from the primary recipients described in ( B ) were pooled for secondary transplantation to sublethally irradiated NSGS mice. Human engraftment was determined by Flow cytometry. Results are means ± SEM of three independent experiments ( n = 8 per group). V, Vehicle; AraC, AraC; QC: Quinacrine; C, CASIN; AraC + QC, AraC + Quinacrine; AraC + QC + C, CASIN + AraC + Quinacrine. Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (* p < 0.05; ** p < 0.01).

    Journal: Nature Communications

    Article Title: Quinacrine-CASIN combination overcomes chemoresistance in human acute lymphoid leukemia

    doi: 10.1038/s41467-021-27300-w

    Figure Lengend Snippet: A Schematic presentation of experimental design. B The effect of CASIN + QC + AraC treatment on total human engraftment in transplanted recipients. 1–3 × 10 4 hCB CD34 + cells were transplanted into sublethally irradiated NSGS mice followed by the treatment described in ( A ). Percentages of human cells (hCD45 + ) in the transplanted recipients 1 week and 8 weeks post treatment were determined by flow cytometry. Results are means ± SEM of three independent experiments (Week 1: V, n = 10; AraC, n = 8; QC, n = 10; AraC + QC, n = 8; CASIN + QC + AraC, n = 8; Week 8: V, n = 8; AraC, n = 9; QC, n = 9; AraC + QC, n = 8; CASIN + QC + AraC, n = 8). C , D The effect of CASIN + QC + AraC treatment on human HSPCs. BM cells were aspirated from the transplanted recipients described in ( B ), and subjected to Flow cytometry analysis for HSPCs (hCD45 + CD34 + CD38 − cells; C ) and HPCs (hCD45 + CD34 + CD38 + cells; D ) 1 week and 8 weeks post treatment. Results are means ± SEM of three independent experiments (Week 1: V, n = 10; AraC, n = 8; QC, n = 8; AraC + QC, n = 8; CASIN + QC + AraC, n = 8; Week 8: V, n = 8; AraC, n = 8; QC, n = 9; AraC + QC, n = 8; CASIN + QC + AraC, n = 9). E CASIN + QC + AraC treatment does not affect long-term repopulating capacity of human HSPCs. BM cells from the primary recipients described in ( B ) were pooled for secondary transplantation to sublethally irradiated NSGS mice. Human engraftment was determined by Flow cytometry. Results are means ± SEM of three independent experiments ( n = 8 per group). V, Vehicle; AraC, AraC; QC: Quinacrine; C, CASIN; AraC + QC, AraC + Quinacrine; AraC + QC + C, CASIN + AraC + Quinacrine. Statistics were performed in the indicated groups: two-tailed, paired t test (parametric); p values are indicated in Source Data files (* p < 0.05; ** p < 0.01).

    Article Snippet: For analysis and sorting of ALL and HSPCs derived from human Cord Blood or adult BM, cells were stained with hCD45-PE Cy7 (Clone: H30, Cat: 560915; BD Biosciences, San Jose, CA), mCD45-PerCP Cy5.5 (Clone:30-F11, Cat: 550994; BD Biosciences, San Jose, CA), Lineage-FITC (Lin1, Cat: 340546; BD Biosciences, San Jose, CA), CD34-PE (Clone 581, Cat: 560941, dilution: 1 in 25; BD Biosciences, San Jose, CA), and CD38-APC (Clone HIT2, Cat: 555462; BD Biosciences, San Jose, CA).

    Techniques: Irradiation, Flow Cytometry, Transplantation Assay, Two Tailed Test

    (A) Experimental scheme for GGF and 3GF LDA using AFT024 monolayers. (B) ELDA of GGF and 3GF cells after 5 weeks of AFT024 LTC, with a 95% CI of 1/2740 – 1/5899 (frequency of 1/4020) for 3GF and 1/4834 – 1/11527 (frequency of 1/7465) for GGF. (C) Experimental scheme for intrahepatic transplants of D15 CD49f+ sorted 3GF cells into newborn NSG mice. (D) Representative flow cytometry contour plots of hCD45 chimerism in CB controls and 2 mice that showed engraftment using D15 3GF cells. (E) Representative flow cytometry contour plots showing the multilineage distribution of week 8 hCD45+ cells from CB controls and D15 3GF IH transplants. CD3+ T cells, CD19+ B cells and CD11c+ myeloid cells. PBMC = peripheral blood mononclear cells and were used as a staining control (non-transplanted).

    Journal: FEBS letters

    Article Title: Induction of human hemogenesis in adult fibroblasts by defined factors and hematopoietic co-culture

    doi: 10.1002/1873-3468.13621

    Figure Lengend Snippet: (A) Experimental scheme for GGF and 3GF LDA using AFT024 monolayers. (B) ELDA of GGF and 3GF cells after 5 weeks of AFT024 LTC, with a 95% CI of 1/2740 – 1/5899 (frequency of 1/4020) for 3GF and 1/4834 – 1/11527 (frequency of 1/7465) for GGF. (C) Experimental scheme for intrahepatic transplants of D15 CD49f+ sorted 3GF cells into newborn NSG mice. (D) Representative flow cytometry contour plots of hCD45 chimerism in CB controls and 2 mice that showed engraftment using D15 3GF cells. (E) Representative flow cytometry contour plots showing the multilineage distribution of week 8 hCD45+ cells from CB controls and D15 3GF IH transplants. CD3+ T cells, CD19+ B cells and CD11c+ myeloid cells. PBMC = peripheral blood mononclear cells and were used as a staining control (non-transplanted).

    Article Snippet: To distinguish levels of engraftment cells were stained for mouse PacBlue-mCD45 (30-F11, Biolegend), and PE-Cy7-hCD45 (2D1, ebioscience).

    Techniques: Flow Cytometry, Staining