JCS032102 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Efficacy and specificity of the lgl1 knockdown. (A) Western blotting of 26 hpf embryonic extracts with anti-Xenopus Lgl1 antibody strongly recognizes bands between 100-120 kDa. One band which is strongly reduced in MO^lgl1-utr-injected embryos (marked with an asterisk) is at the predicted size of Lgl1 around 110 kDa. A lower molecular weight non-specific band (NS) was used as loading control. (B) Injection of MO^lgl1-utr causes heart edema, abnormal eye shapes, reduced forebrain size or post-gastrulation lethality by 26 hpf (not shown). All of these phenotypes could efficiently be rescued by co-injection of lgl1 mRNA that is not targeted by the UTR-directed MO. The rescue efficiency demonstrates high specificity of MO^lgl1-utr induced phenotypes.

• Supplemental Figure S2 -

  Fig. S2. Apical clustering of centrosomes and rosette formation is affected by antagonistic activities of Lgl2 and PRKCI. Epithelial organization of PLLP cells was assayed by immunohistochemistry. Shown are reconstructions of confocal images of the PLLP at 36 hpf. (C-F) The number of rosettes (white brackets) is strongly reduced within different backgrounds. (C′,E′) E-Cadherin is widely expressed along all cell membranes. (D,F) Lack of rosette formation corresponds with a failure of centrosomes to arrange apically.