Journal: Scientific Reports

Article Title: Chymase-producing cells of the innate immune system are required for decidual vascular remodeling and fetal growth

doi: 10.1038/srep45106

Figure Lengend Snippet: (A) Real-time RT-PCR of decidual Mcpt5 mRNA at gd10. (B) Western blot analysis (i, relative expression; ii, blot) of decidual Mcpt5 protein (27 kDa) at gd10. β-actin (42 kDa) was used as a protein loading control. For ( A and B) , the horizontal bars represent medians. Statistical differences were analyzed by using Kruskal Wallis followed by Dunns post test (** P < 0.01, *** P < 0.001). (C) Western blot analysis of Mcpt5 protein in MC/9 cells and PCMCs. (D) Immunofluorescence staining of uNKs (green, DBA lectin, i–ii), Mcpt5 + cells (red, DyLight™ 594, iii–iv) and nuclei (blue, DAPI, vii-viii) in implantation sections from Cpa3 +/+ and Cpa3 Cre/+ mice (scale bars = 20 μm). Overlay (v–viii) reveals Mcpt5 expression in DBA + uNKs. Rectangles represent DAPI staining. (E) Flow cytometry analysis of the percentage of Annexin V + cells following 48 h co-culture of uSMCs with MC/9 cells in the presence or absence of chymase inhibitor from Glycine max (Soybean) (STBI). Results from three independent experiments are presented as median + range. Statistical analysis was performed by using Kruskal Wallis followed by Dunns post test (* P < 0.05). gd, gestation day; PCMCs, peritoneal cavity mast cells; uSMCs, uterine smooth muscle cells.

Article Snippet: For Mcpt5/DBA staining, slides were incubated overnight at 4 °C with an Mcpt5 antibody (Proteintech, 1:50 in 10% BSA in TBS), followed by washing and incubation for 2 h at room temperature with DyLight TM 594-conjugated donkey anti-rabbit secondary antibody (BioLegend, clone 4064, 1:500 in 10% BSA in TBS).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Control, Immunofluorescence, Staining, Flow Cytometry, Co-Culture Assay

Journal: Scientific Reports

Article Title: Chymase-producing cells of the innate immune system are required for decidual vascular remodeling and fetal growth

doi: 10.1038/srep45106

Figure Lengend Snippet: SA wall and lumen diameters from 3 to 10 SAs in Mcpt5-Cre − R-DTA + control mice ( n = 6) or Mcpt5-Cre + R-DTA + mice ( n = 7) were measured. Based on the mean value obtained for each mouse, the means for the group were calculated and expressed as wall thickness (A) and the wall-to-lumen ratio (B) . Statistical differences were analyzed with the Mann-Whitney test (** P < 0.01). (C) Representative images of H/E-stained uterine sections from control (i) and Mcpt5-Cre + R-DTA + Mcpt5-deficient mice (ii, scale bars = 50 μm). (D) Birth weights of pups born to pregnant female controls ( n = 66) or Mcpt5-Cre + R-DTA + mice ( n = 38). Individual mice are depicted, with means. Statistical analysis was performed using the unpaired t test (*** P < 0.001). (E) Pup weights from the Mcpt5-Cre − R-DTA + mice were classified as above the 10 th percentile (blue) or growth restricted (red, below the 5 th percentile; pink, between the 5 th and the 10 th percentile). Pups from wild type Mcpt5-Cre + R-DTA + females served as the basis for calculating the percentiles. SA, spiral artery.

Article Snippet: For Mcpt5/DBA staining, slides were incubated overnight at 4 °C with an Mcpt5 antibody (Proteintech, 1:50 in 10% BSA in TBS), followed by washing and incubation for 2 h at room temperature with DyLight TM 594-conjugated donkey anti-rabbit secondary antibody (BioLegend, clone 4064, 1:500 in 10% BSA in TBS).

Techniques: Control, MANN-WHITNEY, Staining