Glycoprotein 130 promotes human blastocyst development in vitro

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Presentation transcript:

Glycoprotein 130 promotes human blastocyst development in vitro Fredwell Hambiliki, M.Sc., Jörg Hanrieder, Ph.D., Jonas Bergquist, Ph.D., Julius Hreinsson, Ph.D., Anneli Stavreus-Evers, Ph.D., Kjell Wånggren, M.D., Ph.D. Fertility and Sterility Volume 99, Issue 6, Pages 1592-1599.e3 (May 2013) DOI: 10.1016/j.fertnstert.2012.12.041 Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Characteristic protein fingerprint spectra of individual embryos obtained with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) analysis. The peaks represent singly charged individual protein mass peaks that allow preliminary assignment of the respective protein species based on the accurate mass value. The protein annotations are based on their uniprot knowledge base entry (www.uniprot.org). Fertility and Sterility 2013 99, 1592-1599.e3DOI: (10.1016/j.fertnstert.2012.12.041) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) spectra of single intact blastocysts from different treatment groups reveal characteristic changes in protein expression profiles. Different protein intensities for common species as well as exclusively present peaks were observed between the treatment groups (arrows). These species served as characteristic identifier masses for the respective treatment groups. Fertility and Sterility 2013 99, 1592-1599.e3DOI: (10.1016/j.fertnstert.2012.12.041) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Immunohistochemical staining of (A) ubiquitin, (B) histone H4, (C) thymosin beta 4, and (D) thymosin beta-10 in human blastocysts. Nuclear staining with 6-diamino-2-phenylindole (DAPI) is shown in blue. Fertility and Sterility 2013 99, 1592-1599.e3DOI: (10.1016/j.fertnstert.2012.12.041) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Statistical investigation of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) data reveals treatment-specific protein regulations. Unbiased analysis of protein peak intensities by means of multivariate, unpaired t-statistics was performed using the significance analysis of microarrays (SAM) software. (A) (upper left) Control versus gp130 shows significantly lower peak intensities (green) for three protein mass peaks including m/z 8772, m/z 9968, and m/z 19938. (upper right, lower left to lower right) Manual inspection of the MS data was performed for verification of corresponding protein intensity changes (arrows: black trace “control” vs. red trace “gp130”) observed in SAM analysis. (B) Control versus LIF+gp130. The SAM analysis revealed significant changes for many protein peaks (green: lower in LIF+gp130 and red: higher in LIF+gp130). Here thymosin beta chains 10 (*) and 4 (arrow) as well as histone H4 and histone H2B were found to be significantly decreased in LIF+gp130 compared with controls. (C and D) Combinatory LIF+gp130 supplementation versus gp130 and LIF treatment, respectively. The SAM analysis revealed significant changes for many protein peaks. An up-regulation of histone H3 was observed for gp130 compared with LIF+gp130 (C). In contrast, a striking increase of ubiquitin-T was observed in LIF+gp130 compared with sole LIF treatment (D). Fertility and Sterility 2013 99, 1592-1599.e3DOI: (10.1016/j.fertnstert.2012.12.041) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Statistical investigation of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) data reveals treatment-specific protein regulations. Unbiased analysis of protein peak intensities by means of multivariate, unpaired t-statistics was performed using the significance analysis of microarrays (SAM) software. (A) (upper left) Control versus gp130 shows significantly lower peak intensities (green) for three protein mass peaks including m/z 8772, m/z 9968, and m/z 19938. (upper right, lower left to lower right) Manual inspection of the MS data was performed for verification of corresponding protein intensity changes (arrows: black trace “control” vs. red trace “gp130”) observed in SAM analysis. (B) Control versus LIF+gp130. The SAM analysis revealed significant changes for many protein peaks (green: lower in LIF+gp130 and red: higher in LIF+gp130). Here thymosin beta chains 10 (*) and 4 (arrow) as well as histone H4 and histone H2B were found to be significantly decreased in LIF+gp130 compared with controls. (C and D) Combinatory LIF+gp130 supplementation versus gp130 and LIF treatment, respectively. The SAM analysis revealed significant changes for many protein peaks. An up-regulation of histone H3 was observed for gp130 compared with LIF+gp130 (C). In contrast, a striking increase of ubiquitin-T was observed in LIF+gp130 compared with sole LIF treatment (D). Fertility and Sterility 2013 99, 1592-1599.e3DOI: (10.1016/j.fertnstert.2012.12.041) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Statistical investigation of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF MS) data reveals treatment-specific protein regulations. Unbiased analysis of protein peak intensities by means of multivariate, unpaired t-statistics was performed using the significance analysis of microarrays (SAM) software. (A) (upper left) Control versus gp130 shows significantly lower peak intensities (green) for three protein mass peaks including m/z 8772, m/z 9968, and m/z 19938. (upper right, lower left to lower right) Manual inspection of the MS data was performed for verification of corresponding protein intensity changes (arrows: black trace “control” vs. red trace “gp130”) observed in SAM analysis. (B) Control versus LIF+gp130. The SAM analysis revealed significant changes for many protein peaks (green: lower in LIF+gp130 and red: higher in LIF+gp130). Here thymosin beta chains 10 (*) and 4 (arrow) as well as histone H4 and histone H2B were found to be significantly decreased in LIF+gp130 compared with controls. (C and D) Combinatory LIF+gp130 supplementation versus gp130 and LIF treatment, respectively. The SAM analysis revealed significant changes for many protein peaks. An up-regulation of histone H3 was observed for gp130 compared with LIF+gp130 (C). In contrast, a striking increase of ubiquitin-T was observed in LIF+gp130 compared with sole LIF treatment (D). Fertility and Sterility 2013 99, 1592-1599.e3DOI: (10.1016/j.fertnstert.2012.12.041) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions