Ab289363

ScaRNA2 and ATR expression in the TMAs was detected with FISH and immunohistochemistry. The procedures for the scaRNA2 probe hybridization were consistent with the above FISH method. After FITC-TSA incubation, the slides were incubated at 4 °C overnight with ATR primary antibody (Abcam, ab289363, 1:50) in a humidified chamber, followed by the corresponding goat anti-rabbit Cy3 secondary antibody for 1 h at 37 °C in the dark. Image information on the tissue slice was scanned using a panoramic slice scanner (3DHISTECH, Hungary). CaseViewer 2.4 software was employed at 1–400 × arbitrary magnification for observation.

Ovaries were fixed in 4% PFA overnight. After wash and dehydration, ovaries were embedded in paraffin, and sectioned at 8-μm intervals and stained with hematoxylin and eosin. For fluorescent detection, after deparaffinization, sections were treated with Tris-EDTA buffer to retrieve antigen and stained with the 1:500 diluted primary antibodies: pATM (pS1981), BD, 560007; pATR (T1989), Abcam, ab289363; DNA-PK (pS2056), Abcam, ab18192.