human bone marrow stromal cells  (ATCC)

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: B. The impact on the Ezrin-CD44-HA axis of NOMO-1 eIF4E overexpression (eIF4E) compared to vector control (A) or of MM6 CRISPR-4E cells relative to MM6 CRISPR-CTRL cells (B). Left panels. Western blots of eIF4E and Ezrin-CD44-HA axis. Corresponding b-Actin shown for loading controls. Right panels . Confocal micrographs (single section through the plane of cells) of the indicated cells stained for HA or CD44 shown in red. DAPI shown in blue. Scale bar = 10µm. C. Western blot of primary specimens from AML patients (with normal or high-eIF4E) and CD34 + from healthy donors showing expression of eIF4E and Ezrin. b-Actin is provided as a loading control. Each lane represents a different individual. D. Adhesion (left) and invasion (right) capacity using HS-5 stromal cells of NOMO-1 and MM6 cell lines as a function of genetic eIF4E overexpression or CRISPR knockdown respectively. Data is presented as fold change relative to their corresponding controls. Each symbol represents an independent experiment performed with replicates. The bar represents the mean with standard deviations and p-values calculated with two tailed paired T test. E. Schematic representation of colonization assay into mesenchymal stromal cell spheroid model mimicking the bone marrow niche. F. Quantification of AML cell colonization capacity relative to their corresponding controls. Each symbol represents an experimental replicate. The bar represents the mean with standard deviations and p-values, Welch’s t test.

Article Snippet: HS-5 cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Over Expression, Plasmid Preparation, Control, CRISPR, Western Blot, Staining, Expressing, Knockdown, Two Tailed Test

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A . Schematic representation of experimental design using CDX mouse models. B. Percent human cells in the bone marrow at the indicated timepoint assessing the AML engraftment capacity for the indicated cell lines, each dot represents an animal. Violin plot representing the median and quartiles, p values calculated with unpaired two tailed t test and Welch’s t test. C. Kaplan–Meier curves comparing overall survival of mice transplanted with the indicated cell lines. Log-rank (Mantel-Cox) test applied. D. invasion capacity through HS-5 stromal cells of the indicated primary AML samples after exposure to ribavirin or vehicle control. Data is presented as fold change relative to vehicle control. Each symbol is an independent experiment performed with replicates. The bar represents the mean with standard deviations and p-values calculated with Welch’s t test (two tailed). E. Pharmacological targeting of eIF4E with Ribavirin reduces leukemia burden and improves overall survival in an AML patient-derived xenograft (PDX) mouse model. Diagram illustrating the treatment regimen (top panel). Kaplan–Meier curves comparing overall survival of PDX mice with the indicated treatments. Log-rank (Mantel-Cox) test applied (bottom).

Article Snippet: HS-5 cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Two Tailed Test, Control, Derivative Assay

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Western blot of total cell lysates from MM6 cells grown in suspension demonstrated knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) compared to RNAi to luciferase (siLUC) used as a negative control. b-Actin is provided as a loading control. Other proteins of the Ezrin-CD44-HA axis are also shown. Quantification for these is shown in with 3-6 biological replicates for each protein. B. Adhesion and invasion capacity of MM6 cells onto/through HS-5 bone marrow stroma. Fold change relative to siLUC is shown. Each symbol represents a biological replicate performed independently with replicates. Bars represent the mean, shown with standard deviations and p-values (one-way ANOVA). C. Western blot of eIF4E and Ezrin immunoprecipitations (IPs) using total cell lysates from MM6 cells in suspension. SN, supernatant after immunoprecipitation, IgG, negative control. Representative of three biological replicates. IPs of total cell lysates from THP-1 cells in suspension are provided in . D. IPs from MM6 cells in suspension using the rRNA antibody Y10b. LC indicates antibody light chain. Representative of three biological replicates. E. RNA immunoprecipitations (RIPs) from MM6 total cell lysates grown in suspension using anti-Ezrin (Ezrin RIP) or anti-eIF4E (eIF4E RIP) antibodies. Data are from RT-qPCR and represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). F. Western blots of eIF4E and Ezrin IPs from the cytoplasmic fractions of MM6 cells in suspension or G . After invasion through HS-5 bone marrow stroma (invaded). Fractionation controls are provided in . H. RIPs from MM6 cytoplasmic fraction from invaded cells using anti-eIF4E (eIF4E RIP) or anti-Ezrin (Ezrin RIP) antibodies. Data are from RT-qPCR represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). I. Count of the number of pseudopods observed in suspension and invaded MM6 cells represented as a fraction relative to the total cells counted. Each symbol represents a biological replicate. Bars represent the mean, shown with standard deviations and p-values (two-way ANOVA). J . Immunofluorescence and confocal microscopy demonstrating eIF4E, Ezrin, CD44 and rRNA are localized to the same pseudopods (white arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bar = 10 µm

Article Snippet: HS-5 cell line was obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Suspension, Knockdown, Luciferase, Negative Control, Control, Immunoprecipitation, Quantitative RT-PCR, Two Tailed Test, Fractionation, Immunofluorescence, Confocal Microscopy

Journal: NPJ Precision Oncology

Article Title: Venetoclax resistance in preclinical KMT2A-rearranged acute lymphoblastic leukemia models is characterized by high inter- and intra-model heterogeneity

doi: 10.1038/s41698-025-01249-1

Figure Lengend Snippet: A Tumor cell proliferation was monitored by longitudinal full body bioluminescence imaging. Representative images of three animals per group. Mouse IDs are given for each animal to allow for allocation of individual animals in ( B , C ). B Bioluminescence signals from dorsal and ventral images were quantified. 5–8 animals per group, each line represents an individual animal. Mice displayed in ( A ) are marked with the respective mouse ID. Imaging was discontinued once technical signal saturation was achieved. Kolmogorov–Smirnov test for each time point. * p < 0.05. C Quantification of GFP + tumor cell frequency in peripheral blood using flow cytometry. 5–8 animals per group, each line represents an individual animal. Mice displayed in ( A ) are marked with the respective mouse ID. Kolmogorov–Smirnov test for each time point. * p < 0.05; ** p < 0.01. D Kaplan–Meier survival analysis. 6–8 animals per group, Log-rank test. ** p < 0.01. E Relative tumor cell doubling times were calculated based on bioluminescence data ( B ; early proliferation) and peripheral blood flow cytometry values ( C ; late proliferation) and compared to time-matched vehicle cohorts. Mean ± SD of 5–8 animals per group, multiple data sets per animal during the exponential growth phase. Kolmogorov–Smirnov test. * p < 0.05; ** p < 0.01; *** p < 0.001. F Determination of blast frequency in blood, bone marrow and spleen by flow cytometry when the mice reached humane endpoints (30% blasts in blood or weak performance status). Mean ± SD of 4-6 animals per group. Welch’s t test. G Isolated VEN-resistant bone marrow cells were spun onto microscopic slides and Pappenheim stained. Representative images of 5–8 mice per group, ×100 magnification.

Article Snippet: SEM or RS4;11 cells were cultivated alone or cocultured with human stromal bone marrow cells (HS-5 cell line, ATCC, Manassas, VI, USA) and incubated with DMSO (control) or VEN for 72 h. Absolute cell counts were assessed by trypan blue staining and subsequent microscopic quantification.

Techniques: Imaging, Flow Cytometry, Isolation, Staining

Journal: NPJ Precision Oncology

Article Title: Venetoclax resistance in preclinical KMT2A-rearranged acute lymphoblastic leukemia models is characterized by high inter- and intra-model heterogeneity

doi: 10.1038/s41698-025-01249-1

Figure Lengend Snippet: A Quantification of CD45 + /CD19 + tumor cell frequency in peripheral blood using flow cytometry. 1–3 animals per group, each line represents an individual animal. B Kaplan–Meier survival analysis. All five PDX models were summarized. 15 animals per group, Log-rank test. * p < 0.05. C Tumor cell doubling times were calculated based on peripheral blood flow cytometry values ( A ). Mean ± SD of 1–3 animals per group, multiple data sets per animal during exponential growth phase. Unpaired t test. D Tumor cell doubling times summarized for all five PDX models. Mean ± SD of 1–3 animals per group, multiple data sets per animal during exponential growth phase. Kolmogorov–Smirnov test. E Determination of blast frequency in blood, bone marrow and spleen by flow cytometry when the mice reached humane endpoints (30% blasts in blood or weak performance status) or study endpoint (120 days post tumor cell injection). Mean ± SD of 1–3 animals per group. Kolmogorov–Smirnov test. F Tumor cell frequency in blood, bone marrow and spleen at experiment termination, summary of all five PDX models. Mean ± SD of 1–3 animals per group. No statistical assessment due to animals lacking VEN resistance. G Isolated VEN-resistant bone marrow cells were spun onto microscopic slides and Pappenheim stained. Representative images of 1–3 mice per group, 100x magnification. No statistic assessment of data from patients #0054 and #0152 derived models due to animals VEN-treated animals lacking resistance establishment.

Article Snippet: SEM or RS4;11 cells were cultivated alone or cocultured with human stromal bone marrow cells (HS-5 cell line, ATCC, Manassas, VI, USA) and incubated with DMSO (control) or VEN for 72 h. Absolute cell counts were assessed by trypan blue staining and subsequent microscopic quantification.

Techniques: Flow Cytometry, Injection, Isolation, Staining, Derivative Assay