(A) Representative Western blot (WB) of AMOT and associated densitometry in human placenta lysates from 5 to 15 weeks of gestation. AMOT protein levels were normalized by Ponceau staining and expressed as fold change relative to 5–9 weeks. **P < 0.01, ***P < 0.001 by nonparametric Mann-Whitney U test (5–9 weeks, n = 9; 10–15 weeks, n = 10). (B) qPCR for AMOT in human placenta from 5 to 15 weeks of gestation. Data are expressed as fold change relative to 5–9 weeks. *P < 0.05 by nonparametric Mann-Whitney U test (5–9 weeks, n = 10; 10–15 weeks, n = 10). (C) qPCR for AMOT in villous syncytiotrophoblast/cytotrophoblast (ST/CT) layer, and extravillous proximal column (PC) and distal column (DC) in first-trimester placental sections obtained via laser capture microdissection. *P < 0.05, **P < 0.01 by 2-tailed unpaired Student’s t test (5–9 weeks, n = 3; 10–15 weeks, n = 4 or 5). (D) Representative images of IHC staining of AMOT in sections of human placenta from 5 to 6 weeks versus 10 to 12 weeks of gestation (5–6 weeks, n = 7; 10–12 weeks, n = 4). Arrows indicate AMOT localization to specific cell types and structures within the placenta (DC, distal column; IC, intermediate column; PC, proximal column; ST, syncytiotrophoblast; CT, cytotrophoblast; EVT, extravillous trophoblast). Original magnification, ×10 and ×40 (left panel) and ×20 and ×40 (right panel). (E) Representative WB of AMOT and associated densitometry in JEG3 cells following exposure to 21% or 3% oxygen for 24 hours. AMOT protein levels were normalized to β-actin (ACTB) and expressed as fold change relative to cells maintained at 21% oxygen. In A and E, lanes were run on the same gel but were noncontiguous. *P < 0.05 by 2-tailed unpaired Student’s t test (n = 3).