Histo clear reagent

Apoptosis induced DNA damage was measured using the ApopTag® Peroxidase In Situ Apoptosis Detection Kit (S7100, EMD Millipore, Temecula, CA) following the manufacturer’s protocol for paraffin-embedded tissue. Testis sections were deparaffinized using Histo-Clear reagent (HS-200, National Diagnostics, Atlanta, GA). Stained slides were scanned using a Perkin Elmer P250 high capacity slide scanner and images were analyzed using FIJI software to count foci^{38 (link)}.

Tumor sections were placed into Eppendorf tubes and deparaffinized using 1 ml of the HistoClear reagent (National Diagnostics, Atlanta GA) and heated for 12 minutes at 50°C. Samples were centrifuged at 800 rpm for 5 minutes at room temperature, and the supernatant was removed. The remaining pellet was washed twice with 1 ml of 100% ethanol, vortexed and centrifuged at room temperature for 2 minutes at maximum speed, and the ethanol was removed carefully without disturbing the pellet. The pellet was dried at 30°C for 30 minutes. Sequential isolation of RNA from FFPE tissue sections was performed using the MagMAX™ FFPE DNA/RNA Ultra Kit (Thermo Fisher Scientific, Waltham MA) following the manufacturer's instructions. RNA was quantified using the Qubit Fluorometer (Thermo Fisher Scientific, Waltham MA). RNA fragment sizes were analyzed using Experion™ Automated Electrophoresis System (Bio-Rad Laboratories Inc., Hercules CA).