Idasanutlin and venetoclax induces a feedback upregulation of CEBPB and the IL-1/TNF-α pathway. (A) The graphs depict the mean log fold changes (2 replicates) of the indicated genes in MOLM13 and OCIAML3 cells upon overnight idasanutlin (Ida) (300 nM) treatment in comparison with untreated (Un) controls as determined by RNA-seq. (B) The volcano plot depicts DE genes in OCIAML2 cells treated with venetoclax (Ven) (100 nM) in comparison with the DMSO control. (C) The immunoblot analysis shows increasing CEBPB levels with increasing dose gradients of Ven and Ida. (D) Immunoblots showing the CEBPB levels in primary AML samples upon treatment with 2 different doses of Ida or Ven (nM) for 48 hours. Ida+ represents either 30 or 100 nM. Ida++ represents either 100 or 300 nM. Ven+ represents either 3 or 10 nM. Ven++ represents either 10 or 30 nM. CB72 represents CB HSPCs isolated from donor number 72. (E) The immunoblot band areas from panel D were quantified using ImageJ. The area ratios (CEBPB/vinculin) were determined and normalized to no cytokine treatment. Significant differences were determined using multiple comparison analysis of variance tests in comparison with the no cytokine treatment group. (F) Representative FACS histograms depict the increased expression of the myeloid differentiation markers CD11b or CD33 in OCIAML2, OCIAMl3, and/or MOLM13 cells upon treatment with a dose gradient of Ven or Ida. (G) Representative flow cytometry histograms showing the expression changes in the indicated protein after treatment with Ven or Ida for 48 hours. (H) Immunoblots of primary patient samples showing changes in the expression of MDM2 and apoptosis-related proteins when treated with 2 different doses of Ida or Ven.

Idasanutlin and venetoclax induces a feedback upregulation of CEBPB and the IL-1/TNF-α pathway. (A) The graphs depict the mean log fold changes (2 replicates) of the indicated genes in MOLM13 and OCIAML3 cells upon overnight idasanutlin (Ida) (300 nM) treatment in comparison with untreated (Un) controls as determined by RNA-seq. (B) The volcano plot depicts DE genes in OCIAML2 cells treated with venetoclax (Ven) (100 nM) in comparison with the DMSO control. (C) The immunoblot analysis shows increasing CEBPB levels with increasing dose gradients of Ven and Ida. (D) Immunoblots showing the CEBPB levels in primary AML samples upon treatment with 2 different doses of Ida or Ven (nM) for 48 hours. Ida+ represents either 30 or 100 nM. Ida++ represents either 100 or 300 nM. Ven+ represents either 3 or 10 nM. Ven++ represents either 10 or 30 nM. CB72 represents CB HSPCs isolated from donor number 72. (E) The immunoblot band areas from panel D were quantified using ImageJ. The area ratios (CEBPB/vinculin) were determined and normalized to no cytokine treatment. Significant differences were determined using multiple comparison analysis of variance tests in comparison with the no cytokine treatment group. (F) Representative FACS histograms depict the increased expression of the myeloid differentiation markers CD11b or CD33 in OCIAML2, OCIAMl3, and/or MOLM13 cells upon treatment with a dose gradient of Ven or Ida. (G) Representative flow cytometry histograms showing the expression changes in the indicated protein after treatment with Ven or Ida for 48 hours. (H) Immunoblots of primary patient samples showing changes in the expression of MDM2 and apoptosis-related proteins when treated with 2 different doses of Ida or Ven.

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