Fig3c pHH3 and TUNEL co-staining in Wdr62 genetrap adult testis

Raw image dataset for pHH3 and TUNEL co-staining on adult Wdr62 genetrap testis sections. pHH3 staining was performed prior to TUNEL. Please see Fig3a and 3b. 1) Dewax slides2) Antigen retrieval: 10mM Sodium citrate pH6, 95C 15 mins.3) Blocking (20% FBS, 0.2% Triton X in PBS) 1hr RT.4) Primary antibody (rabbit anti-pHH3 (Millipore 06-570) 1:400 diluted in blocking buffer, 4C overnight. 5) Secondary antibody (goat anti-rabbit Alexa 488 (Invitrogen A11034) 1:250 diluted in blocking buffer, RT for 1hr.6) Take 100uL out from labelling reagent #2 for negative control. Then add reagent #1 (50uL) into the rest of the labelling reagent #2 (450uL). Incubate at 37C for 1hr. 7) Counterstain with DAPI 5mg/ml (Sigma D3817) 1:10,000 in PBS for 5 mins RT. 8) Mount in Prolong Diamond (Thermo Fisher Scientific). 9) Images were captured using Leica SP8 inverted confocal microscope.