Extended Data Fig. 5: Isoform-specific dependency on PI3Kγ for Selinexor-induced AKT activation.
a) Immunoblot depicting protein levels of p110-gamma and p101 in OCI-AML2 cells harboring doxycycline-inducible shRNAs targeting PIK3CG (encoding for p110-gamma) and PIK3R5 (encoding for p101) versus scrambled control. B-actin included as loading control. b) Immunoblot depicting protein levels of phosphorylated AKT at T308 in OCI-AML2 cells harboring doxycycline-inducible shRNAs targeting PIK3CA, PIK3CB and PIK3CD versus scrambled control following treatment with selinexor for 36 hours. B-actin included as loading control. c) Immunoblot depicting protein levels of p110α, p110β and p110δ in OCI-AML2 cells harboring doxycycline-inducible shRNAs targeting PIK3CA, PIK3CB and PIK3CD versus scrambled control. B-actin included as loading control. d) Immunoblot depicting protein levels of total and phosphorylated AKT at T308 and S473 in OCI-AML2, MV4;11 and OCI-AML3 cells treated with BYL-719 (PI3K-alpha inhibitor), TGX-221 (PI3K-beta inhibitor), CAL-101 (PI3K-delta inhibitor) or IPI-549 (PI3K-gamma inhibitor) with or without selinexor for 36 hours. OCI-AML2 cells were treated with 500 nM of PI3K inhibitors and MV;411 and OCI-AML3 cells were treated with 100 nM of PI3K inhibitors. B-actin included as loading control. e) Immunoblot depicting protein levels of catalytic PI3K isoforms in OCI-AML2 cells treated with selinexor or DMSO. B-actin included as loading control. f) Immunoblot depicting protein levels of phospho-substrates of PKA or PKC in OCI-AML2 cells treated with selinexor or DMSO. B-actin included as loading control. Representative immunoblots of n = 2-4 biologically independent experiments yielding similar results. B-actin included as loading control.
