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. 2015 Oct 7;10(10):e0140192. doi: 10.1371/journal.pone.0140192

Fig 3. Med28 -/- ICM is not pluripotent.

Fig 3

(A) Semi-quantitative RT-PCR demonstrates reduced expression of pluripotency markers Oct4, Nanog and Sox2 in Med28 -/- (KO) E3.5 blastocysts and cultured blastocysts DIV3 compared to wildtype (WT) controls. Trophoectoderm marker Cdx2 expression is reduced in cultured Med28 -/- (KO) blastocysts. (B) Immunofluorescence analysis shows reduced expression of Oct4 (red, top panels) and Nanog (red, bottom panels) in cultured Med28 -/- blastocysts compared to wildtype controls. (C) Semi-quantitative RT-PCR demonstrates increased expression of primitive endoderm markers Gata4 and Gata6 in DIV3 cultured Med28 -/- (KO) blastocysts, while trophoblast giant cell markers Hand1 and Pl1 expressions are not up-regulated compared to WT control. Gapdh expression serves as a control (shown in A and C). (D) Confocal microscopy images of immunofluorescence analysis show that Gata4 is expressed in primitive endoderm cells surrounding the ICM in DIV3 cultured Med28 +/+ blastocysts (red, top panel) and Med28 -/- blastocysts (red, bottom panel). Nuclear DAPI (blue) and Med28 (green) are also shown (shown in B and D). Scale bar, 100 μm. At least three independent experiments were carried out for all data sets.