Immunohistochemistry (IHC)
The human and rat placental tissue sections were deparaffinized
in xylene and rehydrated through a graded series of ethanol solutions.
Antigen retrieval was performed by immersing the sections in 10 mM
citrate buffer (pH 6.0) and heating them in a pressure cooker for 15
min. Endogenous peroxidase activity was quenched by incubating the
sections in 3% hydrogen peroxide solution for 10 min. Subsequently, the
sections were blocked with 5% inactivated goat serum for 1 h at room
temperature. The sections were then incubated overnight at 4°C with the
following primary antibodies: TP53BP2 (1:5 00, Abcam, Cambridge, USA),
LC3B (1:200, Abcam, Cambridge, USA), and p62 (1:200, Abcam, Cambridge,
USA). After being washed with PBS, the sections were incubated with the
secondary antibody for 2 h at room temperature. Immunoreactivity was
visualized by the addition of 3,3’-diaminobenzidine (DAB) substrate
solution. The sections were counterstained with Harris hematoxylin for
30 s to visualize the cell nuclei. Finally, the sections were dehydrated
through ethanol and xylene, and images were captured via an optical
microscope.