Fig3a pHH3 immunofluorescence in P28 and adult testis

Raw image dataset of phosphohistone H3 (pHH3) immunofluorescence of P28 and adult Wdr62 genetrap testis sections. <br>Please see Shohayeb, B. et al. The association of microcephaly protein WDR62 with CPAP/IFT88 is required for cilia formation and neocortical development. Hum Mol Genet, doi:10.1093/hmg/ddz281 (2019) for protocol.<br>Rehydrated slides containing testis sections were antigen retrieved in 10mM sodium citrate pH6 buffer at 95C for 15mins. Slides were blocked in blocking buffer (20% FBS, 0.2% Triton X in PBS) for at least 1 hour at room temp before incubating with primary antibodies (rabbit anti-phosphohistone H3 (Millipore 06-570) 1:400 diluted in blocking buffer) overnight at 4C. The next day, sections were incubated with secondary antibody (goat anti-rabbit Alexa 488 (Invitrogen A11034) 1:250 and DAPI 5mg/ml (Sigma D8417) 1:10,000 diluted in blocking buffer) for 1hr at room temp. Slides were washed in PBS 3 times before mounting in ProLong Diamond (Thermo Fisher Scientific). Images were captured using Leica SP8 inverted confocal microscope (Leica Microsystems).