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Fig. S2 Morpholino-mediated knockdown of lkb1. A: Western blot analysis of uninjected, spl morpholino- (spl MO), ATG morpholino- (ATG MO), 5 base pair mismatch morpholino- (5 mm MO), and a random 25-mer morpholino-injected (25N MO) samples at 72 hpf. Lkb1 is predicted to have a molecular weight of approximately 50 kD. A second invariant, morpholino-insensitive band is detected at approximately 46 kD. Blots were stripped and probed for α-tubulin as loading control. B: RT-PCR comparing uninjected, spl morphant, 5 mm morphant, and 25N morphant samples at 72 hpf. Samples are normalized to ef1α. C: Western blot analysis of P-RPS6 ser240/244 levels in uninjected, spl MO, and ATG MO samples at 72 hpf, showing induction of phosphorylation in the lkb1 morphant embryos. The blot was stripped and probed with antibody to RPS6 for loading control.