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. 2014 Oct 20;124(11):4941–4952. doi: 10.1172/JCI76864

Figure 7. Effective sFlt1 knockdown in placentas by trophoblast-specific sFLT1 shRNA expression.

Figure 7

(A and B) Blastocysts transduced with lentiviruses expressing GFP control (A) or both sFLT1 shRNA and GFP (B) expressed GFP (denoted by #) in the trophectoderm, but not the inner cell mass. (C and D) Full-thickness placenta on GD18 showing GFP expression in trophoblast lineages from blastocysts transduced with control (C) and sFLT1 shRNA (D) viruses. (E and F) Magnified images from C and D showing GFP expression in the labyrinth layer. (G and H) ISH showing dramatic reduction of sFlt1 signal in sFLT1 shRNA–expressing placentas (H) relative to controls (G); signals were found primarily in GlyTCs and spongiotrophoblasts in the JZ (arrows) and the invading GlyTCs below the giant cell layer (arrowheads). Ch, chorionic plate; La, labyrinth layer. (I) Representative Western blots showing a substantial sFLT1 shRNA–mediated decrease in placental sFLT1 levels on GD18 without changes in VEGF levels. (J–M) qPCR of placental sFlt1 mRNA (J), quantification of placental sFLT1 protein by Western blot (K), and ELISA of maternal serum sFLT1 protein (M) revealed significant reductions at GD18 in placental sFlt1 knockdown animals, whereas (L) placental VEGF/sFLT1 protein ratio was significantly increased by sFlt1 knockdown. Results are mean ± SD. *P < 0.05 (n = 5). Scale bars: 20 μm (A and B); 500 μm (C, D, G, and H); 100 μm (E and F).