Fig. S2.

Role of MMP12 in placental and fetal adaptations to hypoxia. (A) Fetal weights at gd 13.5 from WT and Mmp12 mutant (Δ/Δ607 and Δ/Δ664) rat pregnancies exposed to ambient (Amb) or hypoxia (Hyp) conditions (Ambient, WT: n = 36; Hypoxia, WT: n = 44; Ambient, Δ/Δ607: n = 49; Hypoxia, Δ/Δ607: n = 41; Ambient, Δ/Δ664: n = 35; Hypoxia, Δ/Δ664: n = 41; *P < 0.05). (B) Immunohistochemical analyses (pKRT) of gd 18.5 WT (+/+), Δ/Δ607, and Δ/Δ664 placentation sites. (C) Schematic representation of in vivo maternal exposure to hypoxia (10.5% O₂) and analyses. WT and Mmp12 mutant conceptuses were generated by +/Δ607 male × +/Δ607 female breeding. (D) Effects of hypoxia (10.5% O₂) on the numbers of viable and nonviable conceptuses from WT × WT and heterozygous × heterozygous breeding. Heterozygous × heterozygous pregnancies exposed to hypoxia exhibited a higher number of nonviable conceptuses than did WT × WT (+/+) pregnancies (n = 5/group; *P < 0.05). (E) Immunohistochemical analyses (pan cytokeratin, pKRT; MMP12; elastin) of placentation sites from WT (+/+) and Mmp12 mutant (Δ/Δ607) conceptuses exposed to hypoxic conditions. (F) Quantification of trophoblast cell invasion into the uterine mesometrial compartment (n = 5/group, Mann–Whitney test, *P < 0.05). (G) Genotyping of WT (WT-1) and Mmp12-null (Δ/Δ664–1 and Δ/Δ664–2) TS cells for WT and Δ/Δ664 alleles (Top) and sex chromosome determination of WT and Mmp12-null TS cells (Bottom). WT-1 and Δ/Δ664–2 TS cells are X,Y and Δ/Δ664–1 TS cells are X,X. (H) Conventional RT-PCR analysis of embryonic and trophoblast-specific markers in stem and differentiated WT-1, Δ/Δ664–1, and Δ/Δ664–2 TS cells. (I) Representative images of stem state colonies for WT-1, Δ/Δ664–1, and Δ/Δ664–2 TS cells. Data presented in A and D were analyzed with ANOVA and Student–Newman–Keuls test.