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STEMCELL Technologies Inc rosette sep human monocyte enrichment kit
Rosette Sep Human Monocyte Enrichment Kit, supplied by STEMCELL Technologies Inc, 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/product/rosette-sep+kit/rosettesep+human+monocyte+enrichment+cocktail/pm38535625-251-45-51
Average 90 stars, based on 1 article reviews
rosette sep human monocyte enrichment kit - by Bioz Stars, 2026-10
90/100 stars

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Related Articles

Selection:

Article Title: B-cell reduction using CD37-specific and CD20-specific binding molecules
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit 11 (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Higher Doses of Lenalidomide Are Associated With Unacceptable Toxicity Including Life-Threatening Tumor Flare in Patients With Chronic Lymphocytic Leukemia
Article Snippet: 15 CLL B cells were isolated from freshly donated blood using ficoll density gradient centrifugation (Ficoll-Paque Plus; Amersham Biosciences, Piscataway, NJ) with enrichment using the Rosette-Sep kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: Comparative Assessment of Clinically Utilized CD20-directed Antibodies in Chronic Lymphocytic Leukemia Cells Reveals Divergent NK cell, Monocyte and Macrophage Properties
Article Snippet: Enriched CLL fractions were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: T-cell independent, B-cell receptor-mediated induction of telomerase activity differs among IGHV mutation-based subgroups of chronic lymphocytic leukemia patients
Article Snippet: Isolation of B cells B cells were isolated from PBMCs using the B-cell isolation kit (Miltenyi Biotec) or the Rosette-Sep kit (Stem Cell Technologies) according to the manufacturer's instructions.

Article Title: CD37 immunotherapeutics and uses thereof
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit II (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Arsenic trioxide and Ascorbic Acid Demonstrates Promising Activity against Primary Human CLL Cells in Vitro
Article Snippet: Enriched B-lymphocyte fractions were prepared by using MACS negative selection kit (Miltenyi Biotec, Auburn, CA) or by “Rosette-Sep” kit (Stem Cell Technologies, Vancouver, British Columbia, Canada) according to the manufacturer's instructions.

Article Title: Glycovariant anti-CD37 monospecific protein therapeutic exhibits enhanced effector cell-mediated cytotoxicity against chronic and acute B cell malignancies
Article Snippet: Enriched CLL B cells were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Catolog # 15024) according to the manufacturer's instructions.

Cell Isolation:

Article Title: B-cell reduction using CD37-specific and CD20-specific binding molecules
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit 11 (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Higher Doses of Lenalidomide Are Associated With Unacceptable Toxicity Including Life-Threatening Tumor Flare in Patients With Chronic Lymphocytic Leukemia
Article Snippet: 15 CLL B cells were isolated from freshly donated blood using ficoll density gradient centrifugation (Ficoll-Paque Plus; Amersham Biosciences, Piscataway, NJ) with enrichment using the Rosette-Sep kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: Comparative Assessment of Clinically Utilized CD20-directed Antibodies in Chronic Lymphocytic Leukemia Cells Reveals Divergent NK cell, Monocyte and Macrophage Properties
Article Snippet: Enriched CLL fractions were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: T-cell independent, B-cell receptor-mediated induction of telomerase activity differs among IGHV mutation-based subgroups of chronic lymphocytic leukemia patients
Article Snippet: Isolation of B cells B cells were isolated from PBMCs using the B-cell isolation kit (Miltenyi Biotec) or the Rosette-Sep kit (Stem Cell Technologies) according to the manufacturer's instructions.

Article Title: CD37 immunotherapeutics and uses thereof
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit II (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Arsenic trioxide and Ascorbic Acid Demonstrates Promising Activity against Primary Human CLL Cells in Vitro
Article Snippet: Enriched B-lymphocyte fractions were prepared by using MACS negative selection kit (Miltenyi Biotec, Auburn, CA) or by “Rosette-Sep” kit (Stem Cell Technologies, Vancouver, British Columbia, Canada) according to the manufacturer's instructions.

Article Title: Glycovariant anti-CD37 monospecific protein therapeutic exhibits enhanced effector cell-mediated cytotoxicity against chronic and acute B cell malignancies
Article Snippet: Enriched CLL B cells were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Catolog # 15024) according to the manufacturer's instructions.

Isolation:

Article Title: B-cell reduction using CD37-specific and CD20-specific binding molecules
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit 11 (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Higher Doses of Lenalidomide Are Associated With Unacceptable Toxicity Including Life-Threatening Tumor Flare in Patients With Chronic Lymphocytic Leukemia
Article Snippet: 15 CLL B cells were isolated from freshly donated blood using ficoll density gradient centrifugation (Ficoll-Paque Plus; Amersham Biosciences, Piscataway, NJ) with enrichment using the Rosette-Sep kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: Comparative Assessment of Clinically Utilized CD20-directed Antibodies in Chronic Lymphocytic Leukemia Cells Reveals Divergent NK cell, Monocyte and Macrophage Properties
Article Snippet: Enriched CLL fractions were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: T-cell independent, B-cell receptor-mediated induction of telomerase activity differs among IGHV mutation-based subgroups of chronic lymphocytic leukemia patients
Article Snippet: Isolation of B cells B cells were isolated from PBMCs using the B-cell isolation kit (Miltenyi Biotec) or the Rosette-Sep kit (Stem Cell Technologies) according to the manufacturer's instructions.

Article Title: CD37 immunotherapeutics and uses thereof
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit II (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Arsenic trioxide and Ascorbic Acid Demonstrates Promising Activity against Primary Human CLL Cells in Vitro
Article Snippet: Enriched B-lymphocyte fractions were prepared by using MACS negative selection kit (Miltenyi Biotec, Auburn, CA) or by “Rosette-Sep” kit (Stem Cell Technologies, Vancouver, British Columbia, Canada) according to the manufacturer's instructions.

Article Title: Glycovariant anti-CD37 monospecific protein therapeutic exhibits enhanced effector cell-mediated cytotoxicity against chronic and acute B cell malignancies
Article Snippet: Enriched CLL B cells were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Catolog # 15024) according to the manufacturer's instructions.

Gradient Centrifugation:

Article Title: B-cell reduction using CD37-specific and CD20-specific binding molecules
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit 11 (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Higher Doses of Lenalidomide Are Associated With Unacceptable Toxicity Including Life-Threatening Tumor Flare in Patients With Chronic Lymphocytic Leukemia
Article Snippet: 15 CLL B cells were isolated from freshly donated blood using ficoll density gradient centrifugation (Ficoll-Paque Plus; Amersham Biosciences, Piscataway, NJ) with enrichment using the Rosette-Sep kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: Comparative Assessment of Clinically Utilized CD20-directed Antibodies in Chronic Lymphocytic Leukemia Cells Reveals Divergent NK cell, Monocyte and Macrophage Properties
Article Snippet: Enriched CLL fractions were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: T-cell independent, B-cell receptor-mediated induction of telomerase activity differs among IGHV mutation-based subgroups of chronic lymphocytic leukemia patients
Article Snippet: Isolation of B cells B cells were isolated from PBMCs using the B-cell isolation kit (Miltenyi Biotec) or the Rosette-Sep kit (Stem Cell Technologies) according to the manufacturer's instructions.

Article Title: CD37 immunotherapeutics and uses thereof
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit II (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Arsenic trioxide and Ascorbic Acid Demonstrates Promising Activity against Primary Human CLL Cells in Vitro
Article Snippet: Enriched B-lymphocyte fractions were prepared by using MACS negative selection kit (Miltenyi Biotec, Auburn, CA) or by “Rosette-Sep” kit (Stem Cell Technologies, Vancouver, British Columbia, Canada) according to the manufacturer's instructions.

Article Title: Glycovariant anti-CD37 monospecific protein therapeutic exhibits enhanced effector cell-mediated cytotoxicity against chronic and acute B cell malignancies
Article Snippet: Enriched CLL B cells were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Catolog # 15024) according to the manufacturer's instructions.

Magnetic Cell Separation:

Article Title: B-cell reduction using CD37-specific and CD20-specific binding molecules
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit 11 (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Higher Doses of Lenalidomide Are Associated With Unacceptable Toxicity Including Life-Threatening Tumor Flare in Patients With Chronic Lymphocytic Leukemia
Article Snippet: 15 CLL B cells were isolated from freshly donated blood using ficoll density gradient centrifugation (Ficoll-Paque Plus; Amersham Biosciences, Piscataway, NJ) with enrichment using the Rosette-Sep kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: Comparative Assessment of Clinically Utilized CD20-directed Antibodies in Chronic Lymphocytic Leukemia Cells Reveals Divergent NK cell, Monocyte and Macrophage Properties
Article Snippet: Enriched CLL fractions were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Vancouver, British Columbia, Canada) according to the manufacturer’s instructions.

Article Title: T-cell independent, B-cell receptor-mediated induction of telomerase activity differs among IGHV mutation-based subgroups of chronic lymphocytic leukemia patients
Article Snippet: Isolation of B cells B cells were isolated from PBMCs using the B-cell isolation kit (Miltenyi Biotec) or the Rosette-Sep kit (Stem Cell Technologies) according to the manufacturer's instructions.

Article Title: CD37 immunotherapeutics and uses thereof
Article Snippet: For those samples with less than 90% B cells, negative selection was applied to deplete non-B cells using B cell Isolation Kit II (Miltenyi Biotec, Auburn, Calif.) or by “Rosette-Sep” kit from Stem Cell Technologies (Vancouver, British Columbia, Canada) according to the manufacture suggested protocol.

Article Title: Arsenic trioxide and Ascorbic Acid Demonstrates Promising Activity against Primary Human CLL Cells in Vitro
Article Snippet: Enriched B-lymphocyte fractions were prepared by using MACS negative selection kit (Miltenyi Biotec, Auburn, CA) or by “Rosette-Sep” kit (Stem Cell Technologies, Vancouver, British Columbia, Canada) according to the manufacturer's instructions.

Article Title: Glycovariant anti-CD37 monospecific protein therapeutic exhibits enhanced effector cell-mediated cytotoxicity against chronic and acute B cell malignancies
Article Snippet: Enriched CLL B cells were prepared with the use of the “Rosette-Sep” kit from StemCell Technologies (Catolog # 15024) according to the manufacturer's instructions.



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a , Shown are Gene Set Enrichment Analysis plots for the EPPERT_CE_HSC_LSC (HSC/LSC), EPPERT_PROGENITOR (Progenitor), and HALLMARK_HEME_METABOLISM (Heme Metabolism) gene sets comparing <t>CD34</t> + HSPC with FBXO11 depletion (sh FBXO11 ) to cells expressing non-targeting control shRNA (shCTR). The heatmap on the right is arranged according to pairwise clustering based on the 25 most upregulated and 25 most downregulated genes in FBXO11- depleted cells compared to control. b , Shown are percentages and absolute cell counts of CD34 + HSPC expressing shCTR or sh FBXO11 at day 7 of culture ( n = 4 independent replicates). c , Cell cycle state of CD34 + HSPC was measured by Ki-67/DAPI staining ( n = 3 (shCTR), n = 5 (sh FBXO11 ) independent replicates). d , Percentages of CD45RA - (corresponding to cultured HSC immunophenotype) and CD45RA + 90 - (corresponding to cultured hematopoietic progenitor (HPC) immunophenotype) cells within the CD34 + population are shown ( n = 4 (shCTR), n = 4 (sh FBXO11 ) independent replicates). e , Shown are percentages of megakaryocytic (CD41 + CD61 + ), erythroid (CD71 + Glycophorin A/B + ), and myeloid (CD15 + ) cells in CD34 + HSPC expressing shCTR or sh FBXO11 after 7 days of culture. f , Primary CFC from CD34 + HSPC expressing shCTR or sh FBXO11 were counted 12 days after plating ( n = 5 independent replicates). g , Colony counts following secondary plating of 50,000 cells from the primary CFC assay shown in panel (f) ( n = 5 independent replicates). P values represent two-tailed t -test, and error bars represent s.d.
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Image Search Results


a , Shown are Gene Set Enrichment Analysis plots for the EPPERT_CE_HSC_LSC (HSC/LSC), EPPERT_PROGENITOR (Progenitor), and HALLMARK_HEME_METABOLISM (Heme Metabolism) gene sets comparing CD34 + HSPC with FBXO11 depletion (sh FBXO11 ) to cells expressing non-targeting control shRNA (shCTR). The heatmap on the right is arranged according to pairwise clustering based on the 25 most upregulated and 25 most downregulated genes in FBXO11- depleted cells compared to control. b , Shown are percentages and absolute cell counts of CD34 + HSPC expressing shCTR or sh FBXO11 at day 7 of culture ( n = 4 independent replicates). c , Cell cycle state of CD34 + HSPC was measured by Ki-67/DAPI staining ( n = 3 (shCTR), n = 5 (sh FBXO11 ) independent replicates). d , Percentages of CD45RA - (corresponding to cultured HSC immunophenotype) and CD45RA + 90 - (corresponding to cultured hematopoietic progenitor (HPC) immunophenotype) cells within the CD34 + population are shown ( n = 4 (shCTR), n = 4 (sh FBXO11 ) independent replicates). e , Shown are percentages of megakaryocytic (CD41 + CD61 + ), erythroid (CD71 + Glycophorin A/B + ), and myeloid (CD15 + ) cells in CD34 + HSPC expressing shCTR or sh FBXO11 after 7 days of culture. f , Primary CFC from CD34 + HSPC expressing shCTR or sh FBXO11 were counted 12 days after plating ( n = 5 independent replicates). g , Colony counts following secondary plating of 50,000 cells from the primary CFC assay shown in panel (f) ( n = 5 independent replicates). P values represent two-tailed t -test, and error bars represent s.d.

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: a , Shown are Gene Set Enrichment Analysis plots for the EPPERT_CE_HSC_LSC (HSC/LSC), EPPERT_PROGENITOR (Progenitor), and HALLMARK_HEME_METABOLISM (Heme Metabolism) gene sets comparing CD34 + HSPC with FBXO11 depletion (sh FBXO11 ) to cells expressing non-targeting control shRNA (shCTR). The heatmap on the right is arranged according to pairwise clustering based on the 25 most upregulated and 25 most downregulated genes in FBXO11- depleted cells compared to control. b , Shown are percentages and absolute cell counts of CD34 + HSPC expressing shCTR or sh FBXO11 at day 7 of culture ( n = 4 independent replicates). c , Cell cycle state of CD34 + HSPC was measured by Ki-67/DAPI staining ( n = 3 (shCTR), n = 5 (sh FBXO11 ) independent replicates). d , Percentages of CD45RA - (corresponding to cultured HSC immunophenotype) and CD45RA + 90 - (corresponding to cultured hematopoietic progenitor (HPC) immunophenotype) cells within the CD34 + population are shown ( n = 4 (shCTR), n = 4 (sh FBXO11 ) independent replicates). e , Shown are percentages of megakaryocytic (CD41 + CD61 + ), erythroid (CD71 + Glycophorin A/B + ), and myeloid (CD15 + ) cells in CD34 + HSPC expressing shCTR or sh FBXO11 after 7 days of culture. f , Primary CFC from CD34 + HSPC expressing shCTR or sh FBXO11 were counted 12 days after plating ( n = 5 independent replicates). g , Colony counts following secondary plating of 50,000 cells from the primary CFC assay shown in panel (f) ( n = 5 independent replicates). P values represent two-tailed t -test, and error bars represent s.d.

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: Expressing, shRNA, Staining, Cell Culture, Two Tailed Test

a , Robust multiarray averaging (RMA) normalized FBXO11 signal intensity in CD34 + HSPC samples ( n = 3 independent replicates). b , The Annexin V + population within total (left) or CD34 + (right) cells was analyzed by flow cytometry ( n = 6 (shCTR), n = 7 (sh FBXO11 ) independent replicates). c , Representative plot of cultured CD34 + HSPC analyzed for expression of long-term and short-term HSC (CD90 +/- CD45RA - ) or progenitor (CD90 - CD45RA - ) markers. P values represent two-tailed t -tests and error bars represent s.d. d , Cell cycle states of the HSPC populations were determined by Ki-67 staining ( n = 3 (shCTR), n = 5 (sh FBXO11 ) independent replicates).

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: a , Robust multiarray averaging (RMA) normalized FBXO11 signal intensity in CD34 + HSPC samples ( n = 3 independent replicates). b , The Annexin V + population within total (left) or CD34 + (right) cells was analyzed by flow cytometry ( n = 6 (shCTR), n = 7 (sh FBXO11 ) independent replicates). c , Representative plot of cultured CD34 + HSPC analyzed for expression of long-term and short-term HSC (CD90 +/- CD45RA - ) or progenitor (CD90 - CD45RA - ) markers. P values represent two-tailed t -tests and error bars represent s.d. d , Cell cycle states of the HSPC populations were determined by Ki-67 staining ( n = 3 (shCTR), n = 5 (sh FBXO11 ) independent replicates).

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: Flow Cytometry, Cell Culture, Expressing, Two Tailed Test, Staining

a , Schematic outlining the experimental design for long-term culture and xenotransplant of CD34 + HSPC. b , FBXO11 knockdown maintains CD34 + cells in long-term culture. Percentages (left panel) and total live cell counts (right panel) of CD34 + cells were measured on day 40 of culture ( n = 9 (EV), n = 3 (F, A, FA, K, FK), n = 7 (KA), n = 12 (FKA) independent replicates). c , Kaplan-Meier curve for mice transplanted with cells harvested in (b) ( P = 0.0028 for EV vs FKA, and P = 0.0002 for KA vs FKA (Mantel-Cox test)). d , Bone marrow engraftment of transduced CD34 + cells harvested in (b) (EV, empty vector + control shRNA ( n = 13); KA, KRAS G12D + AML1-ETO ( n = 9); and FKA, sh FBXO11 + KRAS G12D + AML1-ETO ( n = 12) mice) showing CD45 + cell chimerism levels in the non-injected bone (NI) or injected bone (I) ( P = 0.0033 for EV vs FKA, P = 0.018 for KA vs FKA (Sidak’s multiple comparisons test)). e , Giemsa-stained bone marrow smear from mice at endpoint. f , Human CD45 + hematopoietic cell engraftment in the spleen at endpoint as percentage of total viable spleen cells ( n = 5 (EV, 3 not detected (ND)), n = 9 (KA, 1 ND), n = 9 (FKA) mice). g , Human CD45 + hematopoietic cell engraftment in the bone marrow measured at endpoint. ( n = 13 (EV, 7 ND), n = 9 (KA), n = 12 (FKA) mice). h , Percentage of lymphoid (CD3 + , CD19 + ), myeloid (CD33 + ), and other (CD3 - CD19 - CD33 - ) cells within the viable human CD45 + cell population was measured in bone marrow cells at endpoint. ( n = 5 (EV), n = 5 (KA), n = 5 (FKA) mice). i , Total CD34 + HSPC numbers in the bone marrow were quantified at endpoint. ( n = 13 (EV, 13 ND), n = 9 (KA, 8 ND), n = 12 (FKA) mice). j , Hematopoietic cell engraftment in secondary transplants was assessed at weeks 8, 22 and 24. The table on the right lists the fraction of mice in each condition with detectable engraftment at each time point ( P values represent Fisher’s exact tests). All P values represent two-tailed t- tests unless otherwise specified, and error bars represent s.d.

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: a , Schematic outlining the experimental design for long-term culture and xenotransplant of CD34 + HSPC. b , FBXO11 knockdown maintains CD34 + cells in long-term culture. Percentages (left panel) and total live cell counts (right panel) of CD34 + cells were measured on day 40 of culture ( n = 9 (EV), n = 3 (F, A, FA, K, FK), n = 7 (KA), n = 12 (FKA) independent replicates). c , Kaplan-Meier curve for mice transplanted with cells harvested in (b) ( P = 0.0028 for EV vs FKA, and P = 0.0002 for KA vs FKA (Mantel-Cox test)). d , Bone marrow engraftment of transduced CD34 + cells harvested in (b) (EV, empty vector + control shRNA ( n = 13); KA, KRAS G12D + AML1-ETO ( n = 9); and FKA, sh FBXO11 + KRAS G12D + AML1-ETO ( n = 12) mice) showing CD45 + cell chimerism levels in the non-injected bone (NI) or injected bone (I) ( P = 0.0033 for EV vs FKA, P = 0.018 for KA vs FKA (Sidak’s multiple comparisons test)). e , Giemsa-stained bone marrow smear from mice at endpoint. f , Human CD45 + hematopoietic cell engraftment in the spleen at endpoint as percentage of total viable spleen cells ( n = 5 (EV, 3 not detected (ND)), n = 9 (KA, 1 ND), n = 9 (FKA) mice). g , Human CD45 + hematopoietic cell engraftment in the bone marrow measured at endpoint. ( n = 13 (EV, 7 ND), n = 9 (KA), n = 12 (FKA) mice). h , Percentage of lymphoid (CD3 + , CD19 + ), myeloid (CD33 + ), and other (CD3 - CD19 - CD33 - ) cells within the viable human CD45 + cell population was measured in bone marrow cells at endpoint. ( n = 5 (EV), n = 5 (KA), n = 5 (FKA) mice). i , Total CD34 + HSPC numbers in the bone marrow were quantified at endpoint. ( n = 13 (EV, 13 ND), n = 9 (KA, 8 ND), n = 12 (FKA) mice). j , Hematopoietic cell engraftment in secondary transplants was assessed at weeks 8, 22 and 24. The table on the right lists the fraction of mice in each condition with detectable engraftment at each time point ( P values represent Fisher’s exact tests). All P values represent two-tailed t- tests unless otherwise specified, and error bars represent s.d.

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: Plasmid Preparation, shRNA, Injection, Staining, Two Tailed Test

a , Percentages of CD34 + HSPC within the engrafted human hematopoietic cell population at endpoint ( n = 5-12 mice, ND = not detected for values < 0.01%, P values represent Fisher’s exact test). b , Spleen weights were measured at endpoint ( n = 9-13 mice). c , Representative spleen sizes are shown. d , Total live cells harvested from bone marrow at endpoint were counted ( n = 9-13 mice). P values represent two-tailed t- tests unless otherwise specified. Error bars represent s.d..

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: a , Percentages of CD34 + HSPC within the engrafted human hematopoietic cell population at endpoint ( n = 5-12 mice, ND = not detected for values < 0.01%, P values represent Fisher’s exact test). b , Spleen weights were measured at endpoint ( n = 9-13 mice). c , Representative spleen sizes are shown. d , Total live cells harvested from bone marrow at endpoint were counted ( n = 9-13 mice). P values represent two-tailed t- tests unless otherwise specified. Error bars represent s.d..

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: Two Tailed Test

a , Flow cytometry analysis of lymphoid and myeloid cell engraftment at endpoint following xenotransplantion ( n = 5 mice). P values represent two-tailed t- tests and error bars represent s.d. b , Percentages of CD34 + HSPC within the engrafted human hematopoietic cell population at endpoint ( n = 5-12 mice, ND = not detected for values < 0.01%, P values represent Fisher’s exact test). c , Numbers of CD34 + cells generated at xenotransplant endpoint per input CD34 + cell seeded into long-term culture ( n = 9 mice, P values represent two-tailed t -tests). Error bars represent s.d.

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: a , Flow cytometry analysis of lymphoid and myeloid cell engraftment at endpoint following xenotransplantion ( n = 5 mice). P values represent two-tailed t- tests and error bars represent s.d. b , Percentages of CD34 + HSPC within the engrafted human hematopoietic cell population at endpoint ( n = 5-12 mice, ND = not detected for values < 0.01%, P values represent Fisher’s exact test). c , Numbers of CD34 + cells generated at xenotransplant endpoint per input CD34 + cell seeded into long-term culture ( n = 9 mice, P values represent two-tailed t -tests). Error bars represent s.d.

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: Flow Cytometry, Two Tailed Test, Generated

a , Proteins coimmunoprecipitated with FLAG-FBXO11 in K562 cells with or without Benzonase® treatment ( n = 3 to 4 independent replicates). b , Whole cell lysates from K562 parental cells (CTR) or FBXO11 KO CRISPR clones expressing empty vector or FLAG-tagged FBXO11 (FLAG-FBXO11) were immunoblotted as indicated. c , Viable CD34 + HSPC expressing control shRNA (shCTR), sh FBXO11 or sh LONP1 were assessed for megakaryocyte (CD41 + CD61 + ), erythroid (CD71 + CD235a + ) or myeloid (CD14 + or CD15 + ) cell surface markers after 7 days of culture ( n = 4-5 independent replicates). d , MMP measured in K562 and OCI-AML3 cells and CD34 + HSPC expressing shRNA targeting FBXO11 or LONP1 ( n = 3 independent replicates). e , MMP measured in K562 cells expressing WT FBXO11 or FBXO11 with deleted F-box domain (ΔFbox-FBXO11) ( n = 3 independent replicates). f , MMP measured in K562 cells expressing EV with control shRNA (shCTR), or WT-FBXO11 co-expressed with shCTR or shRNA targeting LONP1 (shLONP1). g , Immunoblots of K63-linked polyubiquitination or h , neddylation (NEDD8) of LONP1 immunoprecipitated from parental K562 cells (CTR) or an FBXO11 CRISPR KO clone expressing empty vector or FLAG-FBXO11. i , MMP measured in K562 cells expressing WT-FBXO11 treated with vehicle (DMSO), neddylation activating enzyme inhibitors (MLN4924 or TAS4464), ubiquitin activating enzyme inhibitors (PYR-41 or TAK-243), or j, proteasome inhibitors (MG132 or Bortezomib) ( n = 3 independent replicates). P values represent two-tailed t -tests. Error bars represent s.d.

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: a , Proteins coimmunoprecipitated with FLAG-FBXO11 in K562 cells with or without Benzonase® treatment ( n = 3 to 4 independent replicates). b , Whole cell lysates from K562 parental cells (CTR) or FBXO11 KO CRISPR clones expressing empty vector or FLAG-tagged FBXO11 (FLAG-FBXO11) were immunoblotted as indicated. c , Viable CD34 + HSPC expressing control shRNA (shCTR), sh FBXO11 or sh LONP1 were assessed for megakaryocyte (CD41 + CD61 + ), erythroid (CD71 + CD235a + ) or myeloid (CD14 + or CD15 + ) cell surface markers after 7 days of culture ( n = 4-5 independent replicates). d , MMP measured in K562 and OCI-AML3 cells and CD34 + HSPC expressing shRNA targeting FBXO11 or LONP1 ( n = 3 independent replicates). e , MMP measured in K562 cells expressing WT FBXO11 or FBXO11 with deleted F-box domain (ΔFbox-FBXO11) ( n = 3 independent replicates). f , MMP measured in K562 cells expressing EV with control shRNA (shCTR), or WT-FBXO11 co-expressed with shCTR or shRNA targeting LONP1 (shLONP1). g , Immunoblots of K63-linked polyubiquitination or h , neddylation (NEDD8) of LONP1 immunoprecipitated from parental K562 cells (CTR) or an FBXO11 CRISPR KO clone expressing empty vector or FLAG-FBXO11. i , MMP measured in K562 cells expressing WT-FBXO11 treated with vehicle (DMSO), neddylation activating enzyme inhibitors (MLN4924 or TAS4464), ubiquitin activating enzyme inhibitors (PYR-41 or TAK-243), or j, proteasome inhibitors (MG132 or Bortezomib) ( n = 3 independent replicates). P values represent two-tailed t -tests. Error bars represent s.d.

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: CRISPR, Clone Assay, Expressing, Plasmid Preparation, shRNA, Western Blot, Immunoprecipitation, Two Tailed Test

a , Mitochondrial mass of CD34 + HSPC expressing control shRNA or shRNA targeting FBXO11 or LONP1 was measured using detection of the Mitotracker dye by flow cytometry ( n = 5), P values represent two-tailed t -tests and error bars represent s.d b , Whole cell lysates from K562 parental cells (K562) or FBXO11 KO CRISPR clones (1-20 or 1-48) expressing empty vector (EV) or FLAG-FBXO11 (FBXO11) were immunoblotted for TOM20.

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: a , Mitochondrial mass of CD34 + HSPC expressing control shRNA or shRNA targeting FBXO11 or LONP1 was measured using detection of the Mitotracker dye by flow cytometry ( n = 5), P values represent two-tailed t -tests and error bars represent s.d b , Whole cell lysates from K562 parental cells (K562) or FBXO11 KO CRISPR clones (1-20 or 1-48) expressing empty vector (EV) or FLAG-FBXO11 (FBXO11) were immunoblotted for TOM20.

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: Expressing, shRNA, Flow Cytometry, Two Tailed Test, CRISPR, Clone Assay, Plasmid Preparation

Transcript expression of indicated genes expressed as log 2 (CPM) (counts per million reads mapped) between (b/w) indicated experimental conditions in CD34 + HSPC RNA-seq experiment.

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: Transcript expression of indicated genes expressed as log 2 (CPM) (counts per million reads mapped) between (b/w) indicated experimental conditions in CD34 + HSPC RNA-seq experiment.

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: Expressing, RNA Sequencing Assay

RNA-seq data from CD34 + HSPC expressing shCTR, sh FBXO11 , sh LONP1 , empty vector (Empty or EV), FBXO11, LONP1, FBXO11 with sh LONP1 , or LONP1 with sh FBXO11 ( n = 3 independent replicates except for LONP1 + sh FBXO11 ( n = 2 independent replicates)). a , Heatmap represents pair-wise Euclidian distances between samples based on expression of all detected genes. b , GSEA enrichment plots for the EPPERT_CE_HSC_LSC gene set is shown for various comparisons as noted. c , Enrichment trends of various GSEA genesets are summarized for the comparisons listed. Normalized enrichment scores (NES) for each condition are plotted. The false discovery rate (FDR) is represented by the color scale. d-f , Cell radar plots were generated using human normal hematopoiesis (HemaExplorer) data from the Bloodspot database. The enrichment of genes associated with different hematopoietic populations indicated are compared between different groups as listed. The closer the outline is to the edge of the plot at a labelled point, the greater the proportion of differentially expressed genes associated with the corresponding cell population. Thresholds for differential gene expression were set at log 2 fold > |1.2| and adjusted P < 0.05. d , Shown are proportions of genes associated with various hematopoietic populations upregulated with FBXO11 or LONP1 knockdown compared to control shRNA. e , Shown are proportions of genes associated with various hematopoietic populations downregulated with WT FBXO11 expression, or with FBXO11 expression combined with LONP1 knockdown, relative to empty vector control. f , Shown are proportions of genes associated with various hematopoietic populations down regulated with FBXO11 knockdown, or with FBXO11 knockdown combined with LONP1 expression, relative to control shRNA.

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: RNA-seq data from CD34 + HSPC expressing shCTR, sh FBXO11 , sh LONP1 , empty vector (Empty or EV), FBXO11, LONP1, FBXO11 with sh LONP1 , or LONP1 with sh FBXO11 ( n = 3 independent replicates except for LONP1 + sh FBXO11 ( n = 2 independent replicates)). a , Heatmap represents pair-wise Euclidian distances between samples based on expression of all detected genes. b , GSEA enrichment plots for the EPPERT_CE_HSC_LSC gene set is shown for various comparisons as noted. c , Enrichment trends of various GSEA genesets are summarized for the comparisons listed. Normalized enrichment scores (NES) for each condition are plotted. The false discovery rate (FDR) is represented by the color scale. d-f , Cell radar plots were generated using human normal hematopoiesis (HemaExplorer) data from the Bloodspot database. The enrichment of genes associated with different hematopoietic populations indicated are compared between different groups as listed. The closer the outline is to the edge of the plot at a labelled point, the greater the proportion of differentially expressed genes associated with the corresponding cell population. Thresholds for differential gene expression were set at log 2 fold > |1.2| and adjusted P < 0.05. d , Shown are proportions of genes associated with various hematopoietic populations upregulated with FBXO11 or LONP1 knockdown compared to control shRNA. e , Shown are proportions of genes associated with various hematopoietic populations downregulated with WT FBXO11 expression, or with FBXO11 expression combined with LONP1 knockdown, relative to empty vector control. f , Shown are proportions of genes associated with various hematopoietic populations down regulated with FBXO11 knockdown, or with FBXO11 knockdown combined with LONP1 expression, relative to control shRNA.

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: RNA Sequencing Assay, Expressing, Plasmid Preparation, Generated, shRNA

PCA projection of CD34 + HSPC RNA-seq samples based on top 500 genes with the most variance between samples.

Journal: bioRxiv

Article Title: Loss of FBXO11 function establishes a stem cell program in acute myeloid leukemia through dysregulation of the mitochondrial protease LONP1

doi: 10.1101/2022.09.10.507366

Figure Lengend Snippet: PCA projection of CD34 + HSPC RNA-seq samples based on top 500 genes with the most variance between samples.

Article Snippet: CD34 + HSPCs were obtained at >95% purity from pooled collections using a two-step Rosette-Sep/EasySep human CD34-positive selection kit (STEMCELL Technologies) according to the manufacturer’s protocols and/or FACS sorting.

Techniques: RNA Sequencing Assay