Immunohistochemical Analysis of Paraffin-Embedded Tissues

Paraffin-embedded tissues were sectioned at 4 μm for IHC analysis. Antigen retrieval was performed by incubating the samples in citrate buffer (pH 6.0) for 15 min. After blocking with a mixture of methanol and 0.75% hydrogen peroxide, sections were incubated overnight with primary antibody (VASP, Signalway Antibody, 1:100; POLQ, Signalway Antibody, 1:50), followed by incubation with a secondary antibody conjugated with horseradish peroxidase (goat anti-rabbit, 1:500, Cell Signaling Technology). Sections were washed three times with phosphate-buffered saline and incubated with diaminobenzidine. The process of histologic scoring and analysis was as described in our previous study (Zhou et al., 2021 (link)).