Fig. 2 | Signal Transduction and Targeted Therapy

Fig. 2

From: Activation of RAS/MAPK pathway confers MCL-1 mediated acquired resistance to BCL-2 inhibitor venetoclax in acute myeloid leukemia

Fig. 2

MCL-1 became the primary antiapoptotic protein in VEN-RE cells. a Heatmap showing results of RNA sequencing analysis of BCL-2 family genes in parental and VEN-RE AML cell lines (FDR ≤ 0.2). Genes in red had higher expression, and genes in blue had ower expression, in VEN-RE cells. The numbers in the cells represent normalized expression levels to those in parental lines. b Western blots showing BCL-2 family protein expression in the indicated parental (P) and VEN-RE (R) cell lines. Numbers below the bands are quantifications normalized to levels in the parental lines. c Heatmap showing results of BH3 profiling of the indicated parental and resistant AML cell lines when exposed to the indicated peptides. Data were normalized to DMSO (as negative control) and FCCP (as positive control). The colors indicate the loss of mitochondrial membrane potential (MMP). d Co-immunoprecipitation of parental and VEN-RE MOLM-13 cells protein lysates pulled down with anti-BIM and anti-MCL-1 to determine the binding of BIM with BCL-2 and MCL-1. IgG served as a negative control. e Western blots showing inducible MCL-1 expression. MCL-1 knockdown was induced with 100 ng/mL doxycycline for 0–8 h in VEN-RE MOLM-13 and for 0–16 h in VEN-RE OCI-AML2 cells transfected with a control shRNA (shCON) or shMCL-1. f VEN-RE OCI-AML2 and MOLM-13 cells were treated with 0-4 μM of the MCL-1 inhibitor S63845 with (solid lines) or without 1 μM VEN (dotted lines) for 48 h, then cell viability was determined by CellTiter-Glo assay. Luminescence reads were normalized to those of DMSO-treated controls. Data represent mean ± SD from triplicate independent experiments. g Parental, VEN-RE, and VEN-RE OCI-AML2 and MOLM-13 cells transfected with a control shRNA (shCON) or a shRNA targeting MCL-1 (shMCL-1) that were induced with (solid lines) or without 100 ng/mL doxycycline (dotted lines) and treated with 0–10 μM VEN for 48 h. Cell viability was determined by CellTiter-Glo assay. Luminescence reads were normalized to DMSO-treated control cells. Data are mean ± SD

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