Figure 5.
S63845/MEK inhibitor combination induced CMML monocyte apoptosis. (A-B) Sorted monocytes from healthy donor (gray) or patients with CMML (colors) were treated with 10 nM S63845, 100 nM AZD6244 (AZD, selumetinib) (A), 10 nM trametinib (TRAM) (B), or their combination for 24 hours before measuring the surviving fraction of cells (AnV–/PI–) by flow cytometry (results are normalized to cells treated with dimethyl sulfoxide [DMSO] alone). (C) Immunoblot analysis of indicated proteins in monocytes from patients with CMML treated for 24 hours as in panels A and B. (D) Quantification of the p-ERK/ERK and MCL1/ACTIN ratio using imageQuant LAS 4000 camera and ImageJ software on indicated numbers of immunoblots. (E) Immunoblot analysis of indicated proteins in monocytes from 4 patients with CMML treated for 4 hours as in panels A and B (using imageQuant LAS 4000 camera and ImageJ software). For multiple sample analyses, data are presented as mean ± SEM. Mann-Whitney U test: *P < .05; **P < .01; ***P < .001; ****P < .0001.

S63845/MEK inhibitor combination induced CMML monocyte apoptosis. (A-B) Sorted monocytes from healthy donor (gray) or patients with CMML (colors) were treated with 10 nM S63845, 100 nM AZD6244 (AZD, selumetinib) (A), 10 nM trametinib (TRAM) (B), or their combination for 24 hours before measuring the surviving fraction of cells (AnV/PI) by flow cytometry (results are normalized to cells treated with dimethyl sulfoxide [DMSO] alone). (C) Immunoblot analysis of indicated proteins in monocytes from patients with CMML treated for 24 hours as in panels A and B. (D) Quantification of the p-ERK/ERK and MCL1/ACTIN ratio using imageQuant LAS 4000 camera and ImageJ software on indicated numbers of immunoblots. (E) Immunoblot analysis of indicated proteins in monocytes from 4 patients with CMML treated for 4 hours as in panels A and B (using imageQuant LAS 4000 camera and ImageJ software). For multiple sample analyses, data are presented as mean ± SEM. Mann-Whitney U test: *P < .05; **P < .01; ***P < .001; ****P < .0001.

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