a) Immunoblot depicting protein levels of phosphorylated and total PI3K/AKT pathway members following treatment of a panel of AML cell lines with selinexor. Selinexor was dosed at the following concentrations for each cell line: OCI-AML2 (200nM), MOLM-13 (75nM), MV;411 (50nM), HL-60 (300nM), OCI-AML3 (250nM).
b) Immunoblot depicting protein levels of phosphorylated AKT at T308 and S473 in OCI-AML2, OCI-AML3, and MOLM-13 cells treated with Selinexor for indicated duration.
c) Immunoblot depicting protein levels of XPO1 and total and phosphorylated AKT at T308 in OCI-AML2 cells with doxycycline (dox) inducible shRNAs targeting XPO1 versus scrambled shRNA control. Cells were exposed to dox (75ng/mL) for indicated durations prior to collection.
d) Immunoblot depicting protein levels of XPO1, total and phosphorylated AKT at T308 and S473 in OCI-AML2 cells with sgRNAs targeting XPO1 versus non-targeting controls.
Representative immunoblots of n=3-5 biologically independent experiments yielding similar results. B-actin included as loading control.