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Regulation of male germ cell cycle arrest and differentiation by DND1 is modulated by genetic background DEV057000 Supplementary Material Files in this Data Supplement: Supplemental Table S1 - Supplemental Figure S1 – Fig. S1. Neoplasia formation in 129/SvJ mutant testes. E-cadherin (red). 129/SvJ mutant germ cells express ECAD, MVH, NANOG, SOX2 and OCT4 at E13.5 (A-D, arrows). By E16.5, ECAD-positive neoplasias appear inside testis cords and express the pluripotency markers NANOG, SOX2 and OCT4, but not the germ-cell differentiation gene MVH (E-H). At this stage, there are still a few isolated and morphologically normal germ cells that have faint or no MVH expression (E, inset, arrowheads) but express the pluripotent markers NANOG and SOX2 (F,G, arrowheads). By E19.5, these ECAD-positive masses grow into nascent teratomas. Pluripotent gene expression is still detected, restricted to pockets of cells within the tumor (I-L). Scale bars: 50 μm. Supplemental Figure S2 – Fig. S2. Origin and growth of neoplasias in Dnd1Ter/Ter and Dnd1Ter/+ 129/SvJ Oct4-EGFP-positive testes. (A-D) E15.5 Dnd1Ter/Ter mutant testis. ECAD-positive (red) neoplasias begin to form, which are GFP- (green) and NANOG- (blue) positive. Even cells that morphologically resemble wild-type germ cells are NANOG-positive at this stage (unlike wild-type cells). (E-H) E19.5 Dnd1Ter/+ heterozygote with a neoplasia. At this stage, many isolated germ cells are GFP-positive, NANOG-negative, and still look morphologically normal (arrowheads). The ECAD-positive neoplasia has escaped from the testis cords and grown much larger. Expression of GFP and NANOG are restricted to pockets of cells within the growing tumor (arrows). This phenotype resembles the homozygous Dnd1Ter/Ter mutants on a mixed genetic background, where some germ cells retain germ cell identity at later stages (compare with Fig. 1D-F). Supplemental Figure S3 – Fig. S3. The male somatic program is initiated normally in mutant gonads based on