proliferation  (MedChemExpress)

Journal: Cell Communication and Signaling : CCS

Article Title: Activation of c-Myc confers resistance to venetoclax via inhibition of Bim in t(8;21)-positive acute myeloid leukemia

doi: 10.1186/s12964-026-02994-x

Figure Lengend Snippet: c-Myc down-regulation by HHT abrogates inherent resistance to VEN in t(8;21) AML cells. A c-Myc expression at the mRNA levels after 4 h and 8 h of the indicated treatments ( n = 3 technical replicates). B c-Myc expression at the protein levels after 4 h and 8 h of the indicated treatments ( n = 3 technical replicates). C Western blot analysis of c-Myc expression following treatment with VEN and HHT, in presence or absence of QVD. D Kasumi-1 cells were treated with 50 nM HHT with or without 100 µg/mL cycloheximide (CHX, upper panel), or with or without 40 µM MG-132 (lower panel) for the indicated durations, and c-Myc expression were analyzed by western blot analysis. E Kasumi-1 cells were treated with HHT along with CHX (100 µg/mL) in the absence or presence of MG132 for the indicated durations. c-Myc protein level decreased following CHX treatment, and this decrease was reversed by the simultaneous addition of MG132. F Western blotting analyses of c-Myc expression after treatment with VEN and/or AZA at the indicated concentrations. G Western blotting showing increased c-Myc protein levels in t(8;21) AML cells following treatment with increasing concentrations of VEN. H Relative mRNA expression of c-Myc in t(8;21) AML cells following treatment with VEN, compared to vehicle control. RNA was extracted with TRIzol and subjected to real-time RT-qPCR analysis and normalized to GAPDH. I Kasumi-1 and SKNO-1 cells were treated with VEN (200 nM) and/or 10,058-F4 (60 µM for Kasumi-1, 40 µM for SKNO-1) for 24 h. Cell death was determined by Annexin V-FITC/PI staining and flow cytometry analyses ( n = 3 technical replicates). J Knockdown efficiency of c-Myc was determined by western blotting. K Kasumi-1 cells with c-Myc knockdown were treated with VEN for 48 h, and cell viability was then determined by CCK-8 assay. Data represent five technical replicates. L Kasumi-1 cells with c-Myc knockdown were treated with VEN for 48 h and then subjected to Annexin V-FITC/PI staining and flow cytometry analyses. Data represent four technical replicates. M Overexpression efficiency of c-Myc was determined by western blotting. N Kasumi-1 cells with c-Myc overexpression were treated with VEN for 48 h, and cell viability was then determined by CCK-8 assay. Data represent three technical replicates. Data are mean ± s.d. P values were determined using two-way ANOVA ( A and I ), one-way ANOVA followed by Bonferroni’s test ( H ), or unpaired two-tailed Student’s t-tests ( K , L and N )

Article Snippet: HHT (HY-14944), 10,058-F4 (HY-12702), A-1,331,852 (HY-19741) and S63845 (HY-100741) were purchased from MedChemExpress.

Techniques: Expressing, Western Blot, Control, Quantitative RT-PCR, Staining, Flow Cytometry, Knockdown, CCK-8 Assay, Over Expression, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Activation of c-Myc confers resistance to venetoclax via inhibition of Bim in t(8;21)-positive acute myeloid leukemia

doi: 10.1186/s12964-026-02994-x

Figure Lengend Snippet: c-Myc confers VEN resistance in t(8;21) AML through direct transcriptional repression of Bim. A Scatter plot depicting the correlation between BCL2L11 and MYC mRNA expression in AML1/ETO-positive AML patients. Correlation was assessed using Spearman’s rank correlation coefficient (ρ). B and C c-Myc inhibition activates pro-apoptotic Bim proteins. Representative immunoblot analysis of c-Myc and Bim in lysates from Kasumi-1 cells treated with graded concentrations of 10,058-F4 for 24 h ( B ). Western blot analysis of Bim protein levels in Kasumi-1 cells following c-Myc knockdown( C ). D Representative immunoblot analysis of c-Myc and Bim in lysates from parental MV4-11 (MV4-11P), VEN-resistant MV4-11 (MV4-11R), Kasumi-1, and SKNO-1 cells. E - F CUT&RUN analysis of c-Myc binding to BCL2L11(Bim) promoter region. Enriched DNA samples were amplified using specific primers covering different regions of the BCL2L11 promoter. Input served as a positive control, and IgG was used as a negative control. Fold enrichment values were normalized to the IgG control. ( n = 3 technical replicates). G Validation of CUT&RUN assay by agarose gel electrophoresis. Purified DNA from the Input, anti-c-Myc IP, and normal IgG control groups was amplified by PCR, and the resulting products were resolved on an agarose gel. The Input lane served as a positive control, while the IgG lane was used as a negative control to assess non-specific binding. H c-Myc overexpression suppresses the luciferase activity of a Bim promoter reporter in HEK293T cells. I Validation of c-Myc/Bim double-knockdown efficiency by western blotting. Whole-cell lysates from negative control (NC), single-knockdown (shBim2 or shMyc3), and double-knockdown cells were subjected to immunoblot analysis with antibodies against c-Myc and Bim. β-actin was used as a loading control. J Effect of c-Myc/Bim double-knockdown on VEN sensitivity. Negative control (NC), c-Myc knockdown (shMyc3), and c-Myc/Bim double-knockdown cells (shMyc3 + shBim2) were treated with increasing concentrations of VEN for 48 h. Cell viability was assessed using the CCK-8 assay ( n = 3 technical replicates). P values are indicated numerically on the graph: blue values denote comparisons between shMyc and double-knockdown groups; red values denote comparisons between NC and double-knockdown groups. Data are shown as mean ± SD. P values were determined using unpaired two-tailed Student’s t-tests ( A , E , F and H ) and one-way ANOVA followed by Dunnett’s test( J )

Article Snippet: HHT (HY-14944), 10,058-F4 (HY-12702), A-1,331,852 (HY-19741) and S63845 (HY-100741) were purchased from MedChemExpress.

Techniques: Expressing, Inhibition, Western Blot, Knockdown, Binding Assay, Amplification, Positive Control, Negative Control, Control, Biomarker Discovery, Agarose Gel Electrophoresis, Purification, Over Expression, Luciferase, Activity Assay, CCK-8 Assay, Two Tailed Test