Extended Data Fig. 7: The anti-leukemic effect of AKT inhibition combined with Selinexor in cell line models of AML.
a) Bliss synergy analysis 2D plots for a panel of AML cell lines treated with a dilution series of selinexor and MK-2206. Delta scores indicate synergy (red) and antagonism (green) across the drug-dilution matrix. b) Relative MK-2206 GI50 values in OCI-AML2 cells harboring doxycycline (dox)-inducible shRNAs targeting XPO1 versus scrambled shRNA control. Cells treated with dox for 48 hours prior to incubation with MK-2206 drug-dilution series. Relative MK-2206 GI50 value defined as (GI50 MK-2206 -dox) / (GI50 MK-2206 + dox). c) Time-to-progression assay of OCI-AML2 cells treated with 200 nM selinexor, 5 μM ipatasertib, or the two drugs in combination. d) Selinexor dose–response curves in OCI-AML2 and MOLM-13 cell lines treated with everolimus (50 nM or 100 nM) in the background. e) Relative selinexor GI50 values in OCI-AML2 cells harboring doxycycline-inducible shRNAs targeting P2RY2 versus scrambled shRNA control. Cells were pre-treated with dox for 48 hours prior to incubation with selinexor drug-dilution series. Relative selinexor sensitivity values defined as (GI50 selinexor-dox) / (GI50 selinexor + dox). f) Relative selinexor GI50 values in OCI-AML2 cells co-treated with 5 μM AR-C or DMSO. g) Relative GI50 values of selinexor in combination with PI3K-α/β/δ/γ-specific inhibitors across a panel of AML cell lines. Relative selinexor GI50 value defined as (GI50 selinexor + PI3K inhibitor) / (GI50 selinexor alone). Background PI3K inhibitors dosed by cell line (OCI-AML2, 1 μM; HL-60, 4 μM; MOLM-13, 2 μM; MV4;11, 2 μM; THP-1, 1 μM). h) Immunoblot depicting protein levels of CASPASE 3 and cleaved CASPASE 3 in OCI-AML2 cells treated with selinexor (200 nM), ipatasertib (5 μM), or the combination. Vinculin shown as control. i) Drug-treated induction of annexin positivity in OCI-AML2 cells, as measured by flow cytometry, relative to baseline annexin positivity elicited with DMSO treatment. j) Immunoblot depicting protein levels of BAX in OCI-AML2 cells expressing hairpins targeting BAX or GFP control. B-actin shown as loading control. k) Bliss synergy landscape for two healthy donor cord blood-derived CD34 + cells treated with selinexor versus ipatasertib across a drug-dilution matrix. Extended Data Fig. 7b-g, i data are presented as mean + /-s.e.m. for n = 3 biologically independent replicates. Extended Data Fig. 7b,e-h,i P values computed using multiple unpaired (two-sided) t-tests. Extended Data Fig. 7h,j Representative immunoblots of n = 2 biologically independent experiments yielding similar results.
