Placental Biomarker Immunofluorescence Analysis

Placental section slides were prepared by the Northern Ireland Biobank (38 (link)) using fresh tissue before being subjected to immunofluorescence staining for FKBPL (Cat. no.: 10060-1-AP; Proteintech, UK), SIRT-1 (Cat. no.: ab110304; Abcam, UK), PlGF (Cat. no.: ab180734, Abcam, UK) and VEGF-R1 (Cat. no.: AF321, R&D systems, USA). Tissue slides were imaged using a Leica DMi8 fluorescence inverted microscope using the same magnification (20x) and exposure. Analysis was performed using Image J software (NIH, US) by selecting six random fields of view per section and measuring the intensity, with the assessor blind to patient group. Protein expression was quantified as previously described (39 (link)).