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EBD-300 enhances the anti-leukemic activity of VEN in AML cell lines. (A) Western blot analysis (n=1) reveals lower ASNS protein levels in deletion 7 cell lines (OCI-AML6 and <t>UCSD-AML1)</t> compared to non-deletion 7 cell lines (MOLM-13 and THP-1). (B) Fluorescence in situ hybridization (FISH) analysis shows that all six OCI-AML6 nuclei have only one red (RELN) and one green (TES) fluorescence signal, indicating potential monosomy of chromosome 7 or a deletion in the 7q region. In contrast, all three MOLM-13 nuclei show two pairs of bright fluorescent spots, confirming the presence of both copies of 7q. (C) The cell lines (n=3, technical replicates) were treated with VEH, VEN, or VEN+EBD-300 (0.5 IU/mL) for 96 hours after that Cell titer glo (CTG) assay was used to evaluate cell death. EBD-300 enhances the anti-leukemic effect of VEN in MOLM-13, reducing the VEN IC-50 from 0.0240 µM to 0.0128 µM, and in THP-1, reducing the VEN IC-50 from 5.21 µM to 1.72 µM. OCI-AML6 showed high sensitivity to EBD-300 (IC-50 of 1.1e-4 U/mL), while UCSD-AML1 was resistant to EBD-300. However, EBD-300 still enhanced the anti-leukemic effect of VEN in UCSD-AML1, reducing the VEN IC-50 from 4.34 µM to 2.11 µM. The error bars shown represent the Standard Error of mean or (SEM). *= <0.05, **= <0.01, ***= <0.001, ****= <0.0001.
Ucsd Aml1, supplied by DSMZ, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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EBD-300 enhances the anti-leukemic activity of VEN in AML cell lines. (A) Western blot analysis (n=1) reveals lower ASNS protein levels in deletion 7 cell lines (OCI-AML6 and UCSD-AML1) compared to non-deletion 7 cell lines (MOLM-13 and THP-1). (B) Fluorescence in situ hybridization (FISH) analysis shows that all six OCI-AML6 nuclei have only one red (RELN) and one green (TES) fluorescence signal, indicating potential monosomy of chromosome 7 or a deletion in the 7q region. In contrast, all three MOLM-13 nuclei show two pairs of bright fluorescent spots, confirming the presence of both copies of 7q. (C) The cell lines (n=3, technical replicates) were treated with VEH, VEN, or VEN+EBD-300 (0.5 IU/mL) for 96 hours after that Cell titer glo (CTG) assay was used to evaluate cell death. EBD-300 enhances the anti-leukemic effect of VEN in MOLM-13, reducing the VEN IC-50 from 0.0240 µM to 0.0128 µM, and in THP-1, reducing the VEN IC-50 from 5.21 µM to 1.72 µM. OCI-AML6 showed high sensitivity to EBD-300 (IC-50 of 1.1e-4 U/mL), while UCSD-AML1 was resistant to EBD-300. However, EBD-300 still enhanced the anti-leukemic effect of VEN in UCSD-AML1, reducing the VEN IC-50 from 4.34 µM to 2.11 µM. The error bars shown represent the Standard Error of mean or (SEM). *= <0.05, **= <0.01, ***= <0.001, ****= <0.0001.

Journal: Frontiers in Oncology

Article Title: A mammalian, glutaminase-free asparaginase enhances venetoclax activity in preclinical AML models with chromosome 7 deletion

doi: 10.3389/fonc.2025.1606239

Figure Lengend Snippet: EBD-300 enhances the anti-leukemic activity of VEN in AML cell lines. (A) Western blot analysis (n=1) reveals lower ASNS protein levels in deletion 7 cell lines (OCI-AML6 and UCSD-AML1) compared to non-deletion 7 cell lines (MOLM-13 and THP-1). (B) Fluorescence in situ hybridization (FISH) analysis shows that all six OCI-AML6 nuclei have only one red (RELN) and one green (TES) fluorescence signal, indicating potential monosomy of chromosome 7 or a deletion in the 7q region. In contrast, all three MOLM-13 nuclei show two pairs of bright fluorescent spots, confirming the presence of both copies of 7q. (C) The cell lines (n=3, technical replicates) were treated with VEH, VEN, or VEN+EBD-300 (0.5 IU/mL) for 96 hours after that Cell titer glo (CTG) assay was used to evaluate cell death. EBD-300 enhances the anti-leukemic effect of VEN in MOLM-13, reducing the VEN IC-50 from 0.0240 µM to 0.0128 µM, and in THP-1, reducing the VEN IC-50 from 5.21 µM to 1.72 µM. OCI-AML6 showed high sensitivity to EBD-300 (IC-50 of 1.1e-4 U/mL), while UCSD-AML1 was resistant to EBD-300. However, EBD-300 still enhanced the anti-leukemic effect of VEN in UCSD-AML1, reducing the VEN IC-50 from 4.34 µM to 2.11 µM. The error bars shown represent the Standard Error of mean or (SEM). *= <0.05, **= <0.01, ***= <0.001, ****= <0.0001.

Article Snippet: OCI-AML6 and UCSD-AML1 were purchased from DSMZ (Germany).

Techniques: Activity Assay, Western Blot, Fluorescence, In Situ Hybridization, CTG Assay