Sa00006 2

The myometrial tissues were sliced into 5 μm-thick frozen sections, which were then incubated with a primary rabbit antibody against FPR1 (1:200, ab113531, Abcam) and a mouse anti-myeloperoxidase (MPO) antibody (1:200, 66177-1-Ig, Proteintech) overnight at 4°C. Afterwards, the section were washed with PBS, and then treated with the secondary antibodies, Alex Fluor^®488-conjugated anti-rabbit antibody (1:400, SA00006-2, Proteintech) and CY3-conjugated anti-mouse antibody (1:300, SA00009-1, Proteintech) in the dark at RT for 1 h. Nuclei-staining was done by incubating sections in 5 μg/ml 4,6-diamidino-2-phenylindole (DAPI, Sigma) for 10 min, and then the sections were mounted on glass slides for immunofluorescence analysis.
We cultured the myometrial cells on microscope slides placed in the 12-well plates, then fixed them in 4% formaldehyde for 1 h, permeabilized using 0.2% Triton X-100 and then blocked with 1% BSA for 1 h. The cells were then incubated with a primary rabbit antibody against FPR1 (1:200, ab113531, Abcam) or pMLC (1:100, #3671, CST) overnight. Thereafter, the cells were incubated with FITC fluorescently labeled secondary antibody (1:60, SA00003-2, Proteintech) at RT for 1 h. Cell nuclei were labeled with DAPI. Fluorescent images were captured using a confocal fluorescent microscope.