1. Expression of GRIM-19 in missed abortion and possible pathogenesis
Honglei Chen, M.Sc., Xiaohui Deng, Ph.D., Yang Yang, M.Sc., Yanjun Shen, M.Sc., Lan Chao, Ph.D., Yan Wen, M.Sc., Yanyan Sun, M.Sc. 
Fertility and Sterility 
Volume 103, Issue 1, Pages e3 (January 2015)
DOI: /j.fertnstert
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
GRIM-19 protein and mRNA levels in villous samples. (A, B) Western blot and real-time PCR were performed to assess the protein and relative mRNA levels of GRIM-19 in missed-abortion and control villous samples; ∗∗P<.01. Glyceraldehyde-3-phosphate dehydrogenase served as a loading control. (C) Immunohistochemical analysis was performed to determine the location of GRIM-19 in trophoblasts. Representative microscopic images stained with GRIM-19 antibody are shown (original magnification, ×400).
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Apoptosis in villous tissue. Representative photographs of villous tissue of TUNEL analysis. The apoptotic nuclei showing DNA fragmentation are dark brown. (A) Control samples showed sporadically apoptotic cells in cytotrophoblasts. (B) There were significantly more apoptotic cells in syncytiotrophoblasts and extravillous trophoblast cells in the missed-abortion group than in the control group (original magnification, ×400). (C, D) Western blot and real-time PCR were performed to assess the protein and mRNA levels of P53 in the villous tissue of missed abortion and controls; ∗∗P<.01. Glyceraldehyde-3-phosphate dehydrogenase served as loading control. (E) Immunohistochemical analysis was performed to determine the location of P53 in the trophoblast. Representative microscopic images stained with P53 antibody are shown (original magnification, ×400).
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
Angiogenesis in villous tissue. Representative photographs of villous tissue showing MVD. CD34 immunostaining of vasculature in villous tissue to determine MVD. There was significantly less MVD in the villous samples from (B) missed abortions (23.1 ± 15.6) than in (A) controls (49.3 ± 19.6) (original magnification, ×200). Western blot and real-time PCR were performed to assess the protein and relative mRNA levels of STAT3, HIF-1α, and VEGF in the villous tissue of missed abortion and controls; ∗P<.05, ∗∗P<.01. (C, D) Glyceraldehyde-3-phosphate dehydrogenase served as loading control. The Western blot bands were run separately. (E) Immunohistochemical analysis was performed to determine the locations of STAT3, HIF-1α, and VEGF in the trophoblasts. Representative microscopic images stained with STAT3, HIF-1α, and VEGF antibody are shown (original magnification, ×400).
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
GRIM-19 deficiency and apoptosis and angiogenesis in vitro. (A) Representative flow cytometric analyses of mitochondrial membrane potential using JC-1. In the dot plot, only one subset of cells with high ΔΨ was observed in the SC-siRNA group, but two distinctive subpopulations of cells with both high and low ΔΨ were identified in the G19-siRNA group. The shift in JC-1 fluorescence from red to green indicates a collapse of mitochondrial membrane potential. (B) Representative flow cytogram of annexin V binding (abscissa) vs. PI uptake (ordinate) in the GRIM-19 knockdown HTR-8/SVneo cells and control cells. The numbers in the upper left quadrant (Q1), upper right quadrant (Q2), lower left quadrant (Q3), and lower right quadrant (Q4) represent the relative amount of damaged (annexin V−/PI+), necrotic (annexin V+/PI+), and live (annexin V−/PI−), and relative number of apoptotic cells (annexin V+/PI−), respectively. Western blot was performed to assess the protein levels of STAT3 and HIF-1α in the indicated cell lines. Glyceraldehyde-3-phosphate dehydrogenase served as loading control. (C) The Western blot bands were run separately. Time-dependent secretion of VEGF was determined by ELISA analysis in the indicated cell lines. (D) Each bar represents mean ± SD from three independent experiments; ∗P<.05, ∗∗P<.01.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2015 American Society for Reproductive Medicine Terms and Conditions