Fig 2: Different gene expression dynamics during development of ICM and TE lineages.(A–B) Summary of the number of genes differentially expressed from SCDE analysis (A) between lineages within each developmental time point and (B) between developmental time points within each lineage. Due to the low number of ICM and TE cells at the early and late 16 cell stages, these time points were pooled. (C) Principal component analysis using top 100 variable genes across all cells, annotated for the expression level (log10 RPKM) of early TE-associated genes Krt8, Krt18, Lrp2 and Dab2. (D) TE-specific genes identified by single-cell RNA sequencing associated with at least one Tead4 binding site in trophoblast stem cells (Home et al., 2012). Gene association with Tead4 binding sites was defined as in Home et al. – genes overlapping with Tead4 peaks and genes nearest to Tead4 peaks in 5’ and 3’ directions are considered. Core trophoblast genes (Ralston et al., 2010) are also shown (red line). (E) Representative immunofluorescence stainings of control (Tead4m-z+) and Tead4 maternal/zygotic mutant (Tead4m-z-) embryos for Krt8 and Krt18 (16 cell stage embryos) and Lrp2 and Dab2 (32 to 64 cell stage embryos; Lrp2 and Dab2 were not detected in earlier stage embryos). n indicates total number of embryos analyzed. Scale bar: 25 µm.DOI: http://dx.doi.org/10.7554/eLife.22906.00710.7554/eLife.22906.008Figure 3—source data 1.Excel file of differentially expressed genes from SCDE analysis between lineages within each developmental time point.DOI: http://dx.doi.org/10.7554/eLife.22906.00810.7554/eLife.22906.009Figure 3—source data 2.Excel file of differentially expressed genes from SCDE analysis between developmental time points within each lineage.DOI: http://dx.doi.org/10.7554/eLife.22906.009

Fig 3: ApoJ ? LRP2 signaling is required for insulin-stimulated glucose uptake.a Insulin-stimulated glucose uptake in muscle, adipose tissue, and brown adipose tissue (BAT) was measured in control and L-ApoJ-/- mice (muscle: n = 20 per group, adipose tissue: n = 16 per group, BAT: n = 20 for control, n = 19 for L-ApoJ-/-). Mice were studied at 18 weeks of age. b Insulin-stimulated glucose uptake in muscle, adipose tissue, and BAT was measured in control M-LRP2-/- mice (muscle: n = 10 for control, n = 8 for M-LRP2-/-, adipose tissue: n = 10 for control, n = 7 for M-LRP2-/-, BAT: n = 10 for control, n = 8 for M-LRP2-/-). Mice were studied at 17 weeks of age. Mice were injected with insulin in combination with [14C]2-deoxyglucose. In vivo 2-deoxyglucose uptake was measured. All bars and errors represent means ± SEM. p values by two-sided Student’s t-test in a and b are indicated.

Fig 4: Serum ApoJ is elevated in insulin-resistant subjects with PCOS and normalized by pioglitazone, an insulin-sensitizing drug.a Serum ApoJ levels in normal cycling (normal) and PCOS subjects. Immunoblots represent serum ApoJ and Apo B48 protein in normal and PCOS subjects. Ponceau S staining indicates that equal amounts of protein in each lane was loaded on the gel. Scatter diagram shows serum ApoJ levels measured by ELISA from normal (n = 5) and PCOS subjects (n = 19). b Changes in HOMA2-IR after placebo (n = 8) or pioglitazone treatment (n = 10) in PCOS subjects. c Changes in glucose disposal rate after placebo (n = 8) or pioglitazone (n = 9) treatment in PCOS subjects. d Changes in serum ApoJ levels after placebo (n = 7) or pioglitazone treatment (n = 10) in PCOS subjects. All bars and errors represent means ± SEM. e Relationship of serum ApoJ with fasting insulin levels (n = 38), HOMA2-IR (n = 38), HOMA-ß (%) (n = 38), GDR (n = 40), HDL cholesterol (n = 31), LDL cholesterol (n = 31), free fatty acid (n = 31), and adiponectin (n = 40) levels in normal and PCOS subjects. p values were obtained by Spearman’s rank correlation analysis and r values indicate Spearman’s correlation coefficient. f Muscle ApoJ levels in normal (n = 8) and PCOS subjects (n = 21). PCOS subjects were treated with placebo or pioglitazone. g Muscle LRP2 levels in normal (n = 6) and PCOS subjects (n = 19). h Changes in muscle ApoJ levels after placebo (n = 6) or pioglitazone treatment (n = 12) in PCOS subjects. i Changes in muscle LRP2 levels after placebo (n = 6) or pioglitazone (n = 6) treatment in PCOS subjects. All bars and errors represent means ± SEM. p values by two-sided Student’s t-test in a, c, d, h, and i are indicated.

Fig 5: A role of the ApoJ ? LRP2 signaling pathway in glucose homeostasis and insulin signaling.Circulating ApoJ is dominantly produced by the liver. Liver-derived ApoJ then transports to skeletal muscle where it binds to its receptor LRP2 on the cell surface, leading to increased molecular interaction between LRP2 and IR. The complexes of LRP2 with the IR then undergo co-endocytosis, which is an essential step of insulin signaling in skeletal muscle. As a result, downstream signaling components activate, leading to enhanced glucose transport system activity. Thus, the ApoJ ? LRP2 signaling pathway is central in regulation of glucose homeostasis and insulin signaling.