Fig. 5.

Expression of CD24 and CD47 was down-regulated by ZBTB28. A, B qRT-PCR and Western blot results of CD24 and CD47 expression in stable transfected breast cancer cells, respectively. C Representative images of CD24 and CD47 immunostaining in MB231 xenografts sections of vector and overexpression of ZBTB28. (D) Locations of ChIP-PCR primers (Fragement1 (− 690 ~ -465), 2 (− 486 ~ -323), 3 (− 342 ~ − 122), 4 (− 143 ~  + 18), and 5 (− 2 ~  + 198)) at the CD24 promoter, transcription start site (TSS) was designated as nucleotide + 1. (E) Locations of ChIP-PCR primers (Fragement1 (− 954 ~ − 783), 2 (− 804 ~ − 620), 3 (− 638 ~ − 489), 4 (− 510 ~ − 334), 5 (− 351 ~ − 95), and 6 (− 112 ~  + 66)) at the CD47 promoter, TSS was designated as nucleotide + 1. F, G The effect of ZBTB28 on CD24 and CD47 promoter activity was detected by dual-luciferase reporter system in 293 T, MCF7 and MB231 cells. H, I %Input of CD24 DNA or CD47 DNA by anti-HA antibody was determined by ChIP-PCR. Then, products of ChIP-PCR were used for electrophoresis