Fig. 6

In vivo interaction analysis of AP-1 recognition sequences from the human miR-21 promoter. DNA–protein complexes from SiHa, HeLa, C-33A and HaCaT cells were established using formaldehyde and DNA was fragmented by sonication. DNA was recovered and immunoprecipitations were carried out using anti-c-Fos antibody. PCR amplification products of 475 bp DNA fragment that contains the AP1D, AP1M and AP1P sequences (3 AP1DMP), of 392 bp DNA fragment that contains AP1M and AP1P sequences (2 AP1MP), and of 302 bp DNA fragment that contains the AP1P sequence (1 AP1P) from miR-21 promoter respectively, were resolved in 1% agarose gel electrophoresis using DNA from ChIP assay to SiHa (a), HeLa (b), C-33A (c) and HaCaT cells (d). The DNA 100 bp ladder (MW; lane 1), input condition (In; lane 2), negative control of PCR reaction (C-, lanes 3 and 5), mock condition (M; lane 4), and immunoprecipitation conditions with anti-c-fos antibody (IP; lane 6) were included. The AP-1 sequence from metalloproteinase 1 gene (MMP1) as well as actin housekeeping gene were used as controls respectively. The data are representative at least three independent experiments