DEV02781 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Construction of targeting vector for the generation of Ptf1a-Cre(ex1) mice. (A) The wild-type locus is shown at the top. Filled boxes indicate exons 1 and 2 and open boxes indicate Cre (Cre recombinase) and neo^r (neomycin-resistance gene). The neo^r cassette is flanked by FRT sites (arrowheads; recognition site for Flp recombinase); the transcription start site (ATG) is in the 5′ region of exon 1. The location of the 5′ probe (probe) used for screening ES cells and generated animals by Southern blot analyses is shown. Restriction enzyme sites: E, EcoRI; MI, MluI; Sa, SacI. (B) Southern blot of SacI-digested ES cell genomic DNA. The external 5′ probe detects an 8.5 kb fragment corresponding to the wild-type allele and a 9.5 kb fragment corresponding to the targeted allele. The insertion of the Cre cassette was verified by direct genomic sequencing.

• Supplemental Figure 2 -

  Fig. S2. Localization of β-galactosidase in Ptf1a^+/Cre(ex1) R26R mice during development. (A) Macroscopic view of the dissected abdominal region after X-Gal staining of Ptf1a^+/Cre(ex1) R26R mouse embryo at E18.5. The β-galactosidase expressing tissue (blue) of the pancreas (pa) slightly covers the duodenum (du) and the stomach (st). (B) Whole-mount X-Gal staining (blue) of Ptf1a^+/Cre(ex1) R26R mouse embryo (side view) at E12.5. The black arrow indicates the cerebellum and white arrowheads point to the neural tube. (C) Cross-section of a Ptf1a^+/Cre(ex1) R26R mouse eye at E14.5 showing X-Gal-stained cells in the central retina. (D) X-Gal staining of Ptf1a^+/Cre(ex1) R26R mouse retina at P70. X-Gal-positive cells are visible in the inner region of the neuroretina. (E) Higher magnification of boxed area in C. At E14.5, β-galactosidase-expressing cells are scattered in the inner NBL (arrows, blue). (F) Higher magnification of the boxed area in D. In adult retina, β-galactosidase-expressing cells are localized to the GCL (arrows), central zone of INL and outermost zone of INL (arrowheads). GCL, ganglion cell layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer; rpe, retinal pigment epithelium. Scale bars: 100 μm in C, D; 50 μm in E,F.

• Supplemental Figure 3 -

  Fig. S3. Analysis of cell-cycle status of Ptf1a-expressing cells in wild-type retina. At E13.5 and P1, sections from wild-type mice retinae were stained with antibodies against BrdU (green) and Ptf1a (red). All Ptf1a-expressing cells were negative for the BrdU signal. rpe, retinal pigment epithelium. Scale bar: 50 μm.