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. 2004 Nov 24;101(49):17108–17113. doi: 10.1073/pnas.0407585101

Fig. 5.

Fig. 5.

BRCA1 interacts with MAD2 promoter and positively regulates its function. (A) Luciferase reporter assay of constructs generated by serial deletion. (B) WT and two mutant oligonucleotides from nucleotides 2,131–2,161. The putative OCT1 core like sequence is marked with underlying astral. (C) Biotin-streptavidin pull-down assay. We incubated 1 μg each Biotin labeled oligonucleotide with 300-μg extracts from UBR60 cells 48 h after withdraw of tetracycline. (D) Coimmunoprecipitation assay using antibodies against BRCA1 and OCT1. (C and D) Ten percent of inputs were used in the last lane. (E). Mutation (MT-1) diminished response of MAD2 promoter to BRCA1 induction. (F) Levels of BRCA1 and MAD2 transcripts (Upper) and protein (Lower) in UBR60 cells 48 h after transfection of BRCA1-specific or control RNAi. (G) Depletion of BRCA1 by BRCA1-specific RNAi, but not control RNAi, abolished response of the MAD2 promoter to BRCA1 induction.