Fig. 1

A) Quantification and representative images of SA-b-gal–positive fibroblasts derived from Family I, II, and III after 2 weeks in culture. +/+ genotypes are shown in the chart as pulled together. (B) Immunoblot analysis of p16INK4A protein level in fibroblasts derived from Family I after 2 weeks in culture. (C) Quantification and representative images of the ratio between lens and eye size in control and pik3c2a morphant 72 hours post-fertilization (hpf) zebrafish embryos. (D) Quantification and representative images of SA-b-gal intensity on the lens of control and pik3c2a morphant 72-hpf embryos. (E) Immunoblot analysis of p16INK4A and PI3K-C2α in control and pik3c2a morphant embryos (n = 4 pools of 15 embryos each). (F and G) Confocal images of whole-mount immunofluorescence performed on 72-hpf embryos lens using MKLP1 (red) and Aur-B (green) antibodies to stain midbody and TO-PRO-3 (gray) to stain nuclei. (H) Immunofluorescence of wild-type and Pik3c2a^−/− embryo sections by using a-tubulin to mark intercellular bridges connecting cells in cytokinesis. (I) Quantification of the number of cells connected by bridges (%). n = 6 fields in at least four independent experiments. (J) Time-lapse analysis of the time required to progress from anaphase to cytokinesis in fibroblasts derived from patients with homozygous deletion of PI3K-C2α or in control fibroblasts treated with PITCOIN1. (K) (Left) Immunofluorescence of p16INK4A (red), DNA (blue), and a-tubulin (green) in wild-type and PI3K-C2α-null fibroblast. (Right) Quantification of cell area in control and PIK3C2A-null fibroblasts. If not previously specified, all results are shown as mean or representative picture of at least three independent experiments ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.