Journal: Scientific Reports
Article Title: Healing of Preterm Ruptured Fetal Membranes
Figure Lengend Snippet: Macrophages and adhesion molecules at rupture site. ( A – C ) Immunostaining for F4/80 (green) and DAPI (blue) of membrane 24 h after rupture by 20 G needle. Dashed lines indicate amnion ( Am , white), surface of fetal skin (yellow), chorion ( Cho , light blue), and decidua ( Dec , purple). Myo , myometrium. ( A ) Intact fetal membrane. ( B ) Macrophages within healing amnion (arrows). ( C ) Ruptured edge of chorio-decidua. Bars, 50 µm. ( D – F ) H E staining of ruptured membranes. ( D ) An amniotic fluid macrophage (arrow head) attaches to ruptured amnion at 2 h (arrow). ( E and magnified view in F ) Amniotic fluid macrophages (green) attach to amnion surface at 24 h, and mesenchymal cells migrate (arrow head). Note mesenchymal-like shape change of epithelial cells in F . Bars, 50 µm. ( G and H ) Fluorescent in situ hybridization (FISH) of Y chromosome (red) and immunostaining for F4/80 (green) at healing amnion ( G ) and intact myometrium ( H ) at 48 h. Bars, 10 µm. ( I – L ) Macrophage adhesion molecules at the ruptured site. Immunostaining for F4/80 (green), VCAM1 ( I ) or P-selectin ( J , red) and DAPI (blue) at ruptured amnion at 24 h after rupture by 20 G needle. Bars, 50 µm. ( K and L ) Gene expression of VCAM1 ( K ) or P-selectin ( L ) of intact or ruptured fetal membrane with 20 G needle at indicated time. Error bars represent SEM. n = 5–6 fetal membranes from 5–6 pregnant mice in each group. * P
Article Snippet: Primary antibodies used and concentration were as follows: F4/80 (clone BM8, eBioscience, #14-4801, 1:50), Vimentin (D21H3, Cell Signaling, #5741, 1:100), E-cadherin (4A2, Cell Signaling, #14472, 1:50), VCAM1 (EPR5047, Abcam, #134047, 1:100), P-selectin (CD62P, Abcam, #202983, 1:100), NOS2 (iNOS, Abcam, #15323, 1:100), Arg1 (Arginase-1, D4E3M, Cell Signaling, #93668, 1:100), TNF (Abcam, #6671, 1:100), IL-1β (Abcam, #9722, 1:100), Ly-6G (eBioscience, #14-5931, 1:100), CD68 (KP1, Abcam, #955, 1:100), Ki67 (Abcam #15580, 1:100), HSP70 (Abcam #2787, 1:100).
Techniques: Immunostaining, Staining, In Situ Hybridization, Fluorescence In Situ Hybridization, Expressing, Mouse Assay