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. 2020 Nov 19;5(22):e135775. doi: 10.1172/jci.insight.135775

Figure 4. Genetic modifications in embryonic trophoblast cells.

Figure 4

(A) Morulae were retrieved from pregnant mice (E2.5), grown to blastocysts, infected with lentiviral vectors, and transferred to pseudopregnant mice. Immediately thereafter, Hyal-2 was overexpressed in the trophectoderm. (B) eGFP expression prior to embryo transfer. (C–E) Maximal intensity projections of whole-mount immunofluorescence for Hyal-2 in blastocysts following lentiviral transduction prior to embryo transfer, blastocysts over-expressing HAS-2 in their trophectoderm visualized for eGFP expression, and HAS-2 overexpression following viral infection. (F) Validation of Hyal-2 and HAS-2 overexpression by Western blot analysis. (G) Quantification of Western blot analysis following infection with lentiviral vectors. (H) Histological assessment of hyaluronan deposition as a result of Hyal-2 and HAS-2 overexpression. (n = 3 mice). (I) E6.5 trophoblast cells exclusively expressing eGFP alongside the WT epiblast and endoderm.