Expression levels of cohesin complex factors in ESCs. (A) Mitotic and meiotic cohesin complex model. The mitotic cohesin complex includes SMC1α, RAD21, and STAG1/2. The meiotic cohesin complex contains SMC1β and REC8 instead of the mitotic cohesin components. (B) Scheme of the time course of the experiments. ESCs were synchronized by double thymidine and were released for cell cycle progression. (C) Expression levels of cohesin components in ESCs. After a double thymidine block, the cells were released for cell cycle progression as shown in (B). (D) Expression levels of α-kleisin factors during the S phase. The expression levels of cohesin REC8 and RAD21 were compared during the S phase through western blot analysis and RNA sequencing analysis. (E, F) RNA analysis for cohesin-related genes in undifferentiated and differentiated cells. The transcript levels of the genes involved in cohesin function were analyzed by RNA sequencing (E) and qPCR (F). The primers used in the qPCR analysis were described by Choi et al. (17). The transcript levels were determined by taking the average of two independent analyses from RNA sequencing. After obtaining the qPCR data, the relative expression of the target genes was calculated by using the housekeeping gene 18s rRNA. Three independent experiments were performed, and the error bars represent the mean ± SD.
