Fig. 2: KRAS and RanGAP1 form a protein complex that regulates the release of nuclear protein cargo (EZH2) from the NPC.
a, Complex formation between KRAS and RanGAP1 in A549 cells. Lysates from A549 cells were immunoprecipitated with antibody to RanGAP1 or mock IgG, followed by immunoblotting with antibody to KRAS or RanGAP1. b–e, siRNA knockdown of RanGAP1 (b) abolished the KRAS–NTF2 complex (d; lane 4), whereas siRNA knockdown of NTF2 (c) did not affect the KRAS–RanGAP1 complex (e; lane 4). GAPDH was used as a loading control. f–h, Sotorasib treatment increased the complexes between NUP358 and XPO1 (f and g) and between NUP358 and EZH2 (h). Lysates from sotorasib-treated or sotorasib-untreated NCI-H23 cells were immunoprecipitated with antibody to NUP358 or XPO1 or mock IgG, followed by immunoblotting with antibody to NUP358, XPO1 or EZH2. i–m, Overexpressing mutant KRAS-G12C (i) decreased the complexes between NUP358 and XPO1 (j and k) and between NUP358 and EZH2 (l and m). Lysates from H1703 cells overexpressing mutant KRAS-G12C were immunoprecipitated with antibody to NUP358, XPO1 or EZH2 or mock IgG, followed by immunoblotting with antibody to NUP358, XPO1 or EZH2. Two independent experiments were performed for each image with similar results.
