Fig. 4

p53 directly activates Ell3 transcription. a The accumulation of Ell3 and p53 protein was examined by the immunocytochemical staining of ADSCs transfected with the control or Ell3-expressing plasmid. Scale bar 50 μm. The relative expression of p53 and Ell3 in ADSCs transfected with the control or Ell3-expressing plasmid was analyzed by b qRT-PCR or c immunoblot. d Chromatin immunoprecipitation was performed with control and Ell3 OE cells using antibodies against IgG and p53. The potential p53 binding site (RRRC(A/T)(A/T) GYYY nnnRRRC (A/T) (A/T) GYYY) (− 1113 ~ − 1093) is indicated in red on the ELL3 promoter. e Luciferase assay with the Ell3 promoter. Two hundred ninety-three T cells were transfected with the indicated plasmid for 24 h, and cell lysates were analyzed for luciferase activity. The pGL3-basic plasmid was used as the negative control. f Luciferase assay with deletion mutants of the Ell3 promoter. Deletion mutant-1 contains the promoter region of Ell3 from − 793 to + 493, and deletion mutant-2 contains the promoter region from − 214 to + 493. The experiments were repeated three times independently, and the results presented as bars represent the mean ± s.d. Abbreviations: C, control plasmid; Ell3 OE, Ell3-expressing plasmid; p53 OE, p53-expressing plasmid; WT, Ell3 promoter from − 1260 to + 493; Del #1, Ell3 promoter from − 793 to + 493; Del #2, Ell3 promoter from − 214 to + 493; Luc; luciferase gene