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. 2020 Nov 19;11:587448. doi: 10.3389/fphys.2020.587448

FIGURE 5.

FIGURE 5

Effect of TUNAR on ESCs proliferation and its function during decidualization. (A,B) BrdU cell proliferation assay. The pictures (A) and numbers (B) of the BrdU positive cells were acquired under the fluorescence microscope (100×). BrdU (red) was incorporated into ESCs during S phase. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; blue). (C) PCNA protein levels in ESCs transfected with either TUNAR overexpression vector (OE-TUNAR) or negative control vector (Control); protein samples were collected 48 h after transfection. (D) Expression of TUNAR in ESCs 48 h after transfection with either TUNAR overexpression vector (OE-TUNAR) or negative control vector (Control). The ESCs were treated for 0, 2, 4, 6, 8 days, respectively, then analyzed the mRNA levels of PRL (E), IGFBP1 (F), and level of TUNAR (G) by real-time RT-PCR. Overexpression of TUNAR decreases PRL (H) and IGFBP1 (I) expression in decidualized ESCs. (J) TUNAR expression after transfected with plasmids. Data are representative of at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.