Table 1.
Primers used for realtime PCR, making probes for in situ hybridization, and genotyping of Prss35(−/−) mice.
Figure 1.
Expression of Prss23 in the periimplantation uterus by in situ hybridization.
A. Gestation day 3.5 (D3.5), wild-type (WT, +/+) uterus. B. D3.5 Lpar3(−/−) uterus. C. D4.5 WT uterus. D. D4.5 Lpar3(−/−) uterus. E. D5.5 WT uterus. F. D6.5 WT uterus. G. D7.5 WT uterus. H. D7.5 WT embryo enlarged from the black rectangle in G. I. D7.5 WT uterus enlarged from the red rectangle in G. J. D4.5 WT ovary as a positive control, red arrow indicating granulosa cells. Prss23 sense probe was used as a negative control, no specific signal was detected (data not shown). Red star, embryo; LE, luminal epithelium; Str, stroma; Dec, decidual zone; Myo, myometrium; scale bar, 100 µm.
Figure 2.
Expression of Prss35 in the periimplantation uterus by in situ hybridization.
A. Gestation day 3.5 (D3.5), wild-type (WT, +/+) uterus. B. D3.5 Lpar3(−/−) uterus. C. D4.5 WT uterus. D. D4.5 Lpar3(−/−) uterus. E. D5.5 WT uterus. F. D5.5 Lpar3(−/−) uterus. G. D5.5 WT uterus enlarged from the black rectangle in E. H. D5.5 Lpar3(−/−) uterus enlarged from the black rectangle in F. I. D6.5 WT uterus. M, mesometrial side; AM, antimesometrial side. J. D6.5 WT uterus enlarged from the black rectangle in I. K. D7.5 WT uterus. L. D7.5 WT uterus enlarged from the black rectangle in K. M. D4.5 WT ovary as a positive control. Prss35 sense probe was used as a negative control, no specific signal was detected (data not shown). N. D4.5 Prss35(−/−) ovary. Red star, embryo; LE, luminal epithelium; Str, stroma; Dec, decidual zone; CL, corpus luteum; scale bars, 100 µm.
Figure 3.
Regulation of Prss23 and Prss35 in ovariectomized wild-type mouse uterus upon treatments of progesterone (P4), 17β-estradiol (E2), or P4+E2.
The mRNA expression levels were normalized by the expression of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). HPRT1 (hypoxanthine phosphoribosyltransferase 1) served as the second house-keeping gene. N = 6. Error bars, standard deviation. *p<0.05, compared to vehicle-treated group.
Figure 4.
Deletion of Prss35 on the age of vaginal opening (A) and the number of superovulated cumulus-oocyte complexes (COCs) from each ovary (B).
+/+, wild-type; +/−, Prss35(+/−); −/−, Prss35(−/−); N, the number of female mice (A) or the number of ovaries (B) in each group; error bars, standard deviation.
Figure 5.
Deletion of Prss35 on embryo implantation and the expression of implantation markers in D4.5 uterus.
A. A representative uterus from D4.5 WT mice. B. A representative uterus from D4.5 Prss35(−/−) mice. A & B. Red arrows, implantation sites. C. The average number of implantation sites per mouse in the WT (+/+, N = 8) or Prss35(−/−) (−/−, N = 8) females. All the mating males were Prss35(−/−) for both groups except 2 WT males mated with 2 females in the WT group. Error bars, standard deviation. D∼K. Gene expression in D4.5 uterus by in situ hybridization. D. Prss35 in WT uterus. E. Prss35 in Prss35(−/−) uterus. F. Gjb2 in WT uterus. G. Gjb2 in Prss35(−/−) uterus. H. Gja1 in WT uterus. I. Gja1 in Prss35(−/−) uterus. J. Cox-2 in WT uterus. K. Cox-2 in Prss35(−/−) uterus. Red star, embryo; LE, luminal epithelium; Dec, decidual zone; scale bars, 100 µm.
Figure 6.
Deletion of Prss35 on embryo implantation and the expression of decidualization markers in D7.5 uterus.
A. A representative uterus from D7.5 WT mice. B. A representative uterus from D7.5 Prss35(−/−) mice. A & B. Red arrows, implantation sites. C. The average number of implantation sites per mouse in the WT (+/+, N = 7) or Prss35(−/−) (−/−, N = 7) females. All the mating males were Prss35(−/−) for both groups except one WT male mated with one female in the WT group. Error bars, standard deviation. D∼I. Expression of decidualization markers in D7.5 uterus by in situ hybridization. D. Dtprp in WT uterus. E. Dtprp in Prss35(−/−) uterus. F. Prlpi in WT uterus. G. Prlpi in Prss35(−/−) uterus. H. Abp1 in WT uterus. I. Abp1 in Prss35(−/−) uterus. Red star, embryo; M, mesometrial side; AM, antimesometrial side; scale bars, 500 µm.
Figure 7.
Deletion of Prss35 on litter size. +/+, wild-type; +/−, Prss35(+/−); −/−, Prss35(−/−); N, the number of female mice in each group.
Error bars, standard deviation.
Figure 8.
Deletion of Prss35 on the expression of a few protease-related genes in D4.5 uterus.
+/+, wild-type; −/−, Prss35(−/−). X-axis indicated the names of different genes. Y-axis showed the normalized mRNA expression levels by GAPDH x the number in the parentheses following each gene on X-axis in order to present the relative expression levels of these genes in the same figure. Prss35 was included to confirm that Prss35 was deleted in the Prss35(−/−) uterus. N = 6 (+/+) and N = 4 (−/−). Error bars, standard deviation.