Extended Data Fig. 10. PEN2 and ATP6AP1 control the lysosomal translocation of AXIN.
a, b, Representative images of the experiments shown in Fig. 3g, indicate that PEN2, along with its interaction with ATP6AP1, is required for the lysosomal translocation of AXIN. PEN2-/- MEFs, along with PEN2-/- MEFs re-introduced with wildtype PEN2, its mutant 2A or mutant 20A (a), and ATP6AP1-/- MEFs, along with its wildtype control, or those re-introduced with full length (FL) ATP6AP1, or ATP6AP1Δ420-440 (b, Re-introduced proteins were all expressed at close-to-endogenous levels), were treated with 200 μM metformin for 12 h, or with the v-ATPase inhibitor concanamycin A (conA, 5 μM) for 2 h as a control. AXIN and the lysosomal marker LAMP2 were stained with goat anti-AXIN antibody (green) and rat anti-LAMP2 antibody (red), respectively. Images were taken by confocal microscopy after incubating cells with Alexa Fluor 488 donkey anti-goat IgG and Alexa Fluor 594 donkey anti-rat IgG. The areas defined by dashed boxes on each representative image are enlarged as insets. Experiments in this figure were performed three times.