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Fig. 6 pk1a epilepsy mutants display distinct biochemical properties. (A) Western blot of myc-tagged wild-type (wtPk1) and mutant forms of Pk1a. Protein lysates were made at 80% epiboly stage. β-actin was used as a loading control. (B) Pull-down assays showing the extent to which wild-type (wt) and mutant forms are ubiquitylated. HA-Ub and pk1a RNAs were co-injected into one-cell stage embryos and at 80% epiboly total cell lysates were pulled down by anti-myc antibody beads and blotted for anti-myc or anti-HA. Embryos injected with HA-Ub without pk1a RNA were subjected to anti-myc pull-down as a negative control.