Journal: Cell Reports Medicine

Article Title: PARP-1 improves leukemia outcomes by inducing parthanatos during chemotherapy

doi: 10.1016/j.xcrm.2023.101191

Figure Lengend Snippet:

Article Snippet: Human: HS-5 stromal cells , LGC Standards , ATCC Cat# CRL-11882, RRID: CVCL_3720.

Techniques: Recombinant, Derivative Assay, Staining, Negative Control, Plasmid Preparation, Caspase-3 Assay, TUNEL Assay, Reverse Transcription, Expressing, Bicinchoninic Acid Protein Assay, Western Blot, Microarray, Software, Fluorescence

Journal: Cancer research

Article Title: BONE MARROW STROMA SECRETED CYTOKINES PROTECT JAK2 V617F -MUTATED CELLS FROM THE EFFECTS OF A JAK2 INHIBITOR

doi: 10.1158/0008-5472.CAN-10-4002

Figure Lengend Snippet: (A) FDCP-EpoRV617F cells were cultured alone (control), or with atiprimod, with or without HS5 stromal cells. Following 48 hour culture cells were harvested, stained with annexing V and percentage of apoptotic cells determined by flow cytometry. Data from representative experiment are shown. Atiprimod-induced apoptosis was dramatically impaired when JAK2V617F-positive cells were co-cultured with stromal cells (B) FDCP-EpoRV617F and SET-2 cells were cultured without (control) or with three different stromal cell monolayers (HS5, NK.tert, TM-R1) in the absence or presence of atiprimod for 48 hours. The induction of apoptosis was then assessed by flow cytometry. Results represent the mean ± SD from 3 independent experiments.

Article Snippet: Human stromal cells HS5 (CRL-11882, ATCC, Manassas, VA) were maintained in alpha-MEM medium containing 10% FCS.

Techniques: Cell Culture, Control, Staining, Flow Cytometry

Journal: Cancer research

Article Title: BONE MARROW STROMA SECRETED CYTOKINES PROTECT JAK2 V617F -MUTATED CELLS FROM THE EFFECTS OF A JAK2 INHIBITOR

doi: 10.1158/0008-5472.CAN-10-4002

Figure Lengend Snippet: FDCP-EpoRV617F cells were exposed to 1 μm atiprimod alone (control) or in the presence of HS5 stromal cells (upper panel) or NK.tert stromal cells (lower panel) for 4, 24, and 48 hours. Cells were then lysed and whole cell lysates were immunoprecipitated with a rabbit anti-JAK2 antibody for detection of p-JAK2. Western blot analysis using anti-phosphotyrosine antibody was performed. Then, membranes were stripped and reprobed with anti-JAK2.

Article Snippet: Human stromal cells HS5 (CRL-11882, ATCC, Manassas, VA) were maintained in alpha-MEM medium containing 10% FCS.

Techniques: Control, Immunoprecipitation, Western Blot

Journal: Cancer research

Article Title: BONE MARROW STROMA SECRETED CYTOKINES PROTECT JAK2 V617F -MUTATED CELLS FROM THE EFFECTS OF A JAK2 INHIBITOR

doi: 10.1158/0008-5472.CAN-10-4002

Figure Lengend Snippet: (A) Peripheral blood mononuclear cells from PV patients were cultured alone (control), or with atiprimod, with or without HS5 stromal cells. Following 48 hour culture cells were harvested, stained with annexing V FITC and percentage of apoptotic cells determined by flow cytometry. Data from representative experiment is shown. (B) Ex vivo bone marrow and peripheral blood mononuclear cells from patients with PV were co-cultured with or without HS5, NK.tert, or TM-R1 stromal cells, with or without of 1 μm atiprimod, for 48 hours. Total protein was extracted and subjected to 4-12% NuPAGE® gel and immunoblotting membranes were stained with phospho-STAT3. Membranes were then stripped and reprobed with rabbit anti-total STAT3.

Article Snippet: Human stromal cells HS5 (CRL-11882, ATCC, Manassas, VA) were maintained in alpha-MEM medium containing 10% FCS.

Techniques: Cell Culture, Control, Staining, Flow Cytometry, Ex Vivo, Western Blot

Journal: Cancer research

Article Title: BONE MARROW STROMA SECRETED CYTOKINES PROTECT JAK2 V617F -MUTATED CELLS FROM THE EFFECTS OF A JAK2 INHIBITOR

doi: 10.1158/0008-5472.CAN-10-4002

Figure Lengend Snippet: Human SET2 cells were co-cultured directly (cell on cell) or indirectly (separated by 0.4 μm micropore membranes) with HS5, NK.tert, or TM-R1 stromal cells with or without 1μm atiprimod for 48 hours. Induction of apoptosis was assessed by flow cytometry to compare the direct or indirect effect of stromal monolayers.

Article Snippet: Human stromal cells HS5 (CRL-11882, ATCC, Manassas, VA) were maintained in alpha-MEM medium containing 10% FCS.

Techniques: Cell Culture, Flow Cytometry

Journal: Cancer research

Article Title: BONE MARROW STROMA SECRETED CYTOKINES PROTECT JAK2 V617F -MUTATED CELLS FROM THE EFFECTS OF A JAK2 INHIBITOR

doi: 10.1158/0008-5472.CAN-10-4002

Figure Lengend Snippet: HS5 stromal cells were plated and cultured for 24 hours. Then, stromal cell monolayers were pre-treated with normal goat serum as a control (1 μg), or with goat anti-hIL6, anti-hCXCL10/IP10, and/or anti-hFGF antibodies (0.3 μg), for 4 hours at 37°C. Then, 2×105/mL mouse FDCP-EpoRV617F cells were seeded in the presence of 1μM atiprimod. Control culture without atiprimod has also been done. After 48hrs, JAK2V617F-expressing cells were collected and apoptotic cells were detected by flow cytometry using recombinant human Annexin-V conjugated-APC. Data from representative experiment out of 3 independent experiments is presented; summary results of all 3 experiments are shown too.

Article Snippet: Human stromal cells HS5 (CRL-11882, ATCC, Manassas, VA) were maintained in alpha-MEM medium containing 10% FCS.

Techniques: Cell Culture, Control, Expressing, Flow Cytometry, Recombinant

Journal: Cell chemical biology

Article Title: Allosteric HSP70 inhibitors perturb mitochondrial proteostasis and overcome proteasome inhibitor resistance in multiple myeloma.

doi: 10.1016/j.chembiol.2022.06.010

Figure Lengend Snippet: Figure 2. PI-resistant MM models show increased sensitivity to allosteric HSP70 inhibitors (A and B) AMO1 bortezomib (Btz)-resistant (BtzR) cells are more sensitive than WT cells to an example JG compound, JG194. (C) A larger panel of JG compounds (n = 16) also show increased potency against AMO1-BtzR MM model than WT. (D–F) JG342 exhibits increased potency against MM cell lines resistant to Btz. HSP90 inhibitor 17-DMAG does not show the same phenotype. Bortezomib- sensitive cell lines: KMM1, MM1144, and KMS18. Bortezomib-resistant cell lines: ANBL6-BtzR, LP-1, MMM1, JIM-3, U226-BtzR, RPMI-8226 BtzR, and KMS12BM. (G) JG342 exhibits LC50 values in the nanomolar (nM) range against a panel of MM cell lines (AMO1, AMO1-BtzR, AMO1-CfzR JJN3, KMS11, KMS34, L363, MM.1S, and RPMI8226) and exhibits a therapeutic index against both immortalized (HS5, HS27A) and patient-derived bone marrow stromal lines. All measure- ments in (A)–(G) were performed as n = 4 in 384-well plates, with viability measured using CellTiterGlo at 48 h. (H and I) NSG mice (n = 3 per arm) were implanted with luciferase-labeled RPMI-8226 MM cell line and dosed for 2 weeks with 3 mg/kg JG342 three times per week starting at day 14. JG342 exhibits in vivo anti-MM activity, quantified in (I). All error bars indicate ± SD. p value by two-sided t test.

Article Snippet: HS5 ATCC ATCC Cat#CRL-11882 HS27a ATCC ATCC Cat#CRL-2496 U266 Gift from Brian Van Ness N/A JIM-3 Gift from Brian Van Ness N/A KMS12BM Gift from Brian Van Ness N/A MMM1 Gift from Brian Van Ness N/A LP-1 Gift from Brian Van Ness N/A ANBL6-BtzR Gift from Brian Van Ness N/A U266-BtzR Gift from Brian Van Ness N/A RPMI-8226 BtzR Gift from Brian Van Ness N/A KMM1 Gift from Brian Van Ness N/A KMS18 Gift from Brian Van Ness N/A MM1144 Gift from Brian Van Ness N/A CMK DSMZ DSMZ Cat#ACC 392 Oligonucleotides - See Table S3.

Techniques: Derivative Assay, Luciferase, Labeling, In Vivo, Activity Assay

Journal: PLoS ONE

Article Title: Analysis of acute lymphoblastic leukemia drug sensitivity by changes in impedance via stromal cell adherence

doi: 10.1371/journal.pone.0258140

Figure Lengend Snippet: (A) BLQ5 cells were plated on HS5 stromal cells, resulting in a reduction of impedance (n = 3). (B) Treatment of BLQ5 cells with different concentrations of VX-680 as indicated (n = 3). Dotted lines show the d5 and d9 time points at which cells were harvested for (C) live cell numbers determined by Trypan blue exclusion. Statistical significance was determined using one-way ANOVA, followed by followed by Dunnett’s multiple comparison test. Error bars represent standard deviation. p values relative to DMSO control: ****<0.0001. or (D) Percentage apoptotic cells characterized by FACS using 7-AAD and Annexin V.

Article Snippet: The murine OP9 bone marrow stromal cell line (CRL-2749) and the human HS5 bone marrow stromal cell line (PCS-500-041) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Comparison, Standard Deviation, Control

Journal: PeerJ

Article Title: Integrated analysis of programmed cell death-related genes identifies CORO1A as an apoptosis-associated gene in acute myeloid leukemia

doi: 10.7717/peerj.21303

Figure Lengend Snippet: (A) CORO1A expression levels in HS-5, HL-60 and THP-1 cells. (B) CORO1A expression levels in HL-60 and CORO1A knockdown HL-60 cell lines. (C) CORO1A expression levels in THP-1 and CORO1A knockdown THP-1 cell lines. (D) CCK-8 analysis of CORO1A knockdown HL-60 and THP-1cell lines. (E) Bcl-2 and Bax expression levels in CORO1A knockdown HL-60 cell line. (F) Bcl-2 and Bax expression levels in CORO1A knockdown THP-1 cell line. (G) Annexin V staining analysis of CORO1A knockdown HL-60 cell line. (H) Annexin V staining analysis of CORO1A knockdown THP-1cell line.

Article Snippet: Human HS-5, HL-60, and THP-1 cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Knockdown, CCK-8 Assay, Staining