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. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: Nat Cell Biol. 2015 May 4;17(6):726–735. doi: 10.1038/ncb3168

Figure 5. PICC-1 physical interactions with PAC-1 and JAC-1.

Figure 5

(a) Yeast two-hybrid interactions between PICC-1 fused to the GAL4 DNA-binding domain (DBD-bait) and GAL-4 activation domain fusions with PAC-11-574 or JAC-1 (AD-prey). Yeast was plated on selective medium requiring an interaction for growth (see Methods); strong (‘++’: DBD-Krev1 + AD-RalGDS), medium (‘+’: DBD-Krev1 + AD-RalGDSm1), and no interaction (‘−’: DBD-Krev1 + AD-RalGDSm2) controls are shown for comparison. (b) Immunoprecipitation of JAC-1 (four isoforms are visible) and co-immunoprecipitation of PICC-1-GFP from embryonic lysates; jac-1(xn15) mutants (‘xn15’) expressing PICC-1-GFP were used as a control; ‘WT’ = wild-type jac-1 allele. (c) Immunoprecipitation of PICC-1-GFP and co-immunoprecipitation of mCherry-HA-PAC-1 from embryonic lysates; wild-type embryos, and embryos expressing mCherry-HA-PAC-1 but not PICC-1-GFP were used as controls. Immunoprecipitation experiments shown in (b) and (c) were performed twice, and representative examples are shown. Tubulin levels in the input (total embryonic lysate) are shown as a loading control. ‘IB,’ Immunoblot; ‘IP,’ Immunoprecipitation. Uncropped blots are shown in Supplementary Figure 8.