Fig. 5

CDC25A overexpression can rescue ES cell lethality of Brca2^KO/KO. a Immunoblot showing IR-induced degradation of CDC25A in PL2F7 mES cells expressing either HA-BRE or HA-BRE mutant defective in USP7 binding (FLΔ123). GAPDH was used as control. The histogram below shows the average relative abundance of CDC25A at particular time points from three independent experiments. All intensities were normalized against corresponding GAPDH band intensities. Supplementary Table 2 shows the P values. b Representative Southern blot analysis showing the identification of Brca2^KO/KO cells (marked with solid stars) after CRE-mediated deletion of conditional allele in indicated cell lines. Upper band: conditional allele (CKO); lower band: knock-out allele (KO). c Western blot showing CDC25A abundance in Brca2^CKO/KO and Brca2^CKO/KO; MSCV-Cdc25A ES cells before (lanes 1, 3, 5) and 2 h after 6 Gy IR (lanes 2, 4, 6). Top panel represents the scheme of expression of FLAG-CDC25A from MSCV LTR. GAPDH was used as a loading control. d Southern blot analysis of HAT-resistant ES cell colonies after CRE-mediated deletion of conditional allele in Brca2^CKO/KO; MSCV-Cdc25A ES cells to identify Brca2^KO/KO clones (marked with solid stars), upper band: conditional allele (CKO); lower band: knock-out allele (KO). e CDC25A mRNA expression in two different breast cancer data sets from The Cancer Genome Atlas (TCGA). Left panel represents the data set of METABRIC breast cancer with 2433 tumors and right panel shows the breast invasive carcinoma data set of 825 tumors. Dots or squares in graphs represent individual tumors. Middle line, median; whiskers, minimal and maximum values. Significance was calculated using unpaired t-test with Welch’s corrections