Supplementary Information for
Heaney JD, Anderson EL, Michelson MV, Zechel JL, Conrad PA, Page DC, Nadeau JH
Germ cell pluripotency, premature differentiation and susceptibility to testicular teratomas in mice.
Development 139, 1577 (2012)
- Figure S1 Confocal microscopy images of Oct4::GFP transgenic, E13.5 testes immunostained for KI67 and E15.5 testes immunostained for KI67 and E-cadherin: (A) E13.5 FVB, (B) E13.5 M19, (C) E15.5 FVB and (B) E15.5 M19. KI67-positive, E-cadherin-negative E15.5 germ cells are indicated (arrow). Scale bars: 75 Ám.
- Figure S2 CCND1 expression is downregulated with the initiation of meiosis in female germ cells. Confocal microscopy images of Oct4::GFP transgenic E15.5 female germ cells immunostained for CCND1 and ?H2A.X. (A) CCND1 and ?H2A.X immunostaining are shown for the E15.5 Oct4::GFP transgenic female gonad shown in Fig. 1. Dotted lines indicate the boundary at which germ cells transition from mostly ?H2A.X-positive to mostly CCND1-positive. The anterior (A) to posterior (P) axis is indicated. Scale bar: 150 Ám. (B) Magnified view of CCND1 and ?H2A.X staining in E15.5 female germ cells. CCND1 and ?H2A.X are not expressed in the same cells. Scale bar: 75 Ám.
- Figure S3 Co-expression of CCND1, NANOG and KI67 in EC cell foci at E16.5. (A) Single color panels of CCND1, NANOG, or KI67 and E-cadherin staining in serial sections of an Oct4::GFP transgenic, E16.5 M19 testis (same testis as shown in Fig. 6B). E-cadherin-positive, low GFP-expressing EC cell foci are indicated (arrows). Scale bar: 150 Ám. (B) Confocal microscopy images of serial sections of an E16.5 FVB testis stained for CCND1, NANOG or KI67. Oct4::GFP transgenic germ cells are negative for both CCND1 and NANOG. Scale bar: 75 Ám.
- Figure S4 SYCP3 and SYCP1 localization in germ cells. Chromosome spreads from E14.5 and E16.5 gonads were stained for DAPI and SYCP3 or SYCP1, and imaged by confocal microscopy. (A) Pre-leptotene, nucleolar localization of SYCP3 in E14.5 M19 male germ cells. Scale bar: 100 Ám. (B) Pachytene distribution of SYCP1 on synapsed chromosomes in an E16.5 female germ cell. Teratoma-resistant and susceptible male germ cells were negative for SYCP1 staining (not shown). Scale bar: 10 Ám.