The Immunohistochemistry (IHC) experiments were performed as our previous reported10 (link). Briefly, the paraffin-embedded tumor tissues were cut into 4 μm. The tissues were retrieved in citric acid buffer (PH = 7.0), and then were blocked with normal goat serum (Boster Biological Technology, Wuhan, China) for 60 min at room temperature. The tissues were incubated with primary antibody (anti-Foxp1, cat no. Ab16645, 1:200 dilution, Abcam; anti-Ki67, cat no. Ab92742, 1:500 dilution, Abcam) overnight at 4 °C. The tissues were then then with a horseradish peroxidase (HRP) -conjugated anti-rabbit secondary antibody for 2 h at 37 °C. Finally, the sections were counterstained with hematoxylin.
The expression of Foxp1 and Ki67 was semi-quantitated by immunoreactivity scoring. The intensity of Foxp1 staining was scored as 0 (negative), 1 (weak), 2 (moderate), and 3 (intense) by two pathologists who were blinded to the experiments. The immunoreactivity score was calculated as the percentage of positive cells multiplied by the intensity of staining.
The expression of Foxp1 and Ki67 was semi-quantitated by immunoreactivity scoring. The intensity of Foxp1 staining was scored as 0 (negative), 1 (weak), 2 (moderate), and 3 (intense) by two pathologists who were blinded to the experiments. The immunoreactivity score was calculated as the percentage of positive cells multiplied by the intensity of staining.
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