POU5F1 and central nervous system cancer: OCT4 transcripts were also reduced in CRISPRi-HML-2–transfected cells with RNA in situ hybridization 48 hours after transfection; furthermore, we detected decreased HML-2 env and OCT4 proteins by immunostaining at 72 hours after transfection in CRISPRi-HML-2 glioma neurospheres compared with the ngCRISPRi construct (Figure 7, C and D).