Figure 3.

Lack of fetus-derived IGF2 reduces the expansion of feto-placental microvasculature in late gestation

(A) Functions enriched in DEGs at E19.

(B) qRT-PCR analysis of angiopoietin-Tie2/TEK signaling components in Lz (n = 6–8 per group).

(C) TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) staining in E16 Lz (arrows point to apoptotic cells) and data quantification (n = 6 samples per group); scale bars, 50 μm.

(D) Left: representative double immunostaining for TUNEL (red) and laminin (green, marker of feto-placental capillaries) in the Lz of an E16 Igf2^EpiKO mutant placenta (DAPI, blue marks the nuclei; white and red arrows indicate TUNEL⁺ FPECs and LT, respectively; scale bars, 25 μm). Right: quantification of TUNEL⁺ cells that are positive or negative for laminin (n = 6 Igf2^EpiKO mutant placentae).

(E) Feto-placental endothelial cell (FPEC) proliferation measured by flow cytometry (left—representative histograms at E16; right—data quantification; n = 4–11 per group).

(F) qRT-PCR analysis of Adgre1 in Lz.

(G) Representative F4/80 immunostainings in E16 Lz (arrows indicate macrophages). Scale bars, 100 μm. Right: percentage of macrophages/Lz at E16 (n = 6–8 samples per group).

(H) Representative CD31 immunostaining in Lz (scale bars, 100 μm).

(I) qRT-PCR analysis for SynT-II (syncytiotrophoblast layer II) marker genes. For all graphs, data are presented as averages or individual values; error bars are SD; ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001 by two-way ANOVA plus Sidak’s multiple comparisons tests in (B), (C), (E), (F), and (I) or Mann-Whitney tests in (G). See also Figure S4 and Table S2.