Cryoprotectants up-regulate expression of mouse oocyte AQP7, which facilitates water diffusion during cryopreservation  Ya-Jing Tan, M.Sci., Yun Xiong, B.Sci., Guo-Lian Ding, Ph.D., M.D., Dan Zhang, Ph.D., M.D., Ye Meng, M.D., He-Feng Huang, M.D., Jian-Zhong Sheng, Ph.D., M.D.  Fertility and Sterility  Volume 99, Issue 5, Pages 1428-1435 (April 2013) DOI: 10.1016/j.fertnstert.2012.11.049 Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Expression of AQP7 mRNA was detected in human (a, 283 bp) and mouse (b, 177 bp) oocytes, but not detected in negative control (NC). (B) Immunofluorescence analysis confirmed AQP7 expression in human and mouse oocytes (green: AQP7; blue: nuclear DNA). (C) Photographs of typical oocytes preincubated with (+Hg) or without (−Hg) of 50 μM HgCl2 for 15 minutes before and 60 or 120 seconds after exposure to hypotonic medium. (D–E) Summary data of changes in the cell volume (D) and Pf (E) in control and HgCl2-treated groups. V, oocyte volume at different time. V0, initial oocyte volume. Data are present as mean ± standard error. **P<.01 compared with the corresponding control. n, number of repeated experiments. Bar = 20 μm. Fertility and Sterility 2013 99, 1428-1435DOI: (10.1016/j.fertnstert.2012.11.049) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Immunofluorescence analysis of AQP7 expression in mouse oocytes in the presence of (A–B) 8% EG, (C–D) 9.5% DMSO, and (E–F) 0.5 M sucrose, respectively. Data are presented as mean ± SE. **P<.01 compared with the corresponding controls. n, number of repeated experiments. Bar = 20 μm. Fertility and Sterility 2013 99, 1428-1435DOI: (10.1016/j.fertnstert.2012.11.049) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 3 (A) Expression of AQP7 mRNA in 293T cells tranfected with GFP-hAQP7 fusion protein expression vector. (B–C) Increased green fluorescent protein (GFP) intensities in 293T cells tranfected with GFP-hAQP7 fusion protein expression vector or GFP vector alone in the presence of 8% EG, 9.5% DMSO, or 0.5 M sucrose, respectively (B and C). Data are presented as mean ± SE. n, number of repeated experiments. Bar = 10 μm. Fertility and Sterility 2013 99, 1428-1435DOI: (10.1016/j.fertnstert.2012.11.049) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 4 (A–B) Photographs of typical oocytes and the changes in cell volume of mouse oocytes in the presence of 8% EG and 9.5% DMSO, respectively. V, oocyte volume at different time. V0, initial oocyte volume. t, duration from minimal oocyte volume to 90% of initial volume regained. (C) Survival rate of oocytes treated by vitrificated cryopreservation for 48 hours. Data are presented as mean ± standard error. n, number of repeated experiments. *P<.05 (chi-square test). Bar = 20 μm. Fertility and Sterility 2013 99, 1428-1435DOI: (10.1016/j.fertnstert.2012.11.049) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions