Journal: Cell Death & Disease
Article Title: FK506 binding protein 51 positively regulates melanoma stemness and metastatic potential
doi: 10.1038/cddis.2013.109
Figure Lengend Snippet: FKBP51 interacts with the general transcriptional co-activator p300. ( a , left) FKBP51 co-immunoprecipitates with p300. ( a , right), p300 co-immunoprecipitates with FKBP51. Total-cell lysates were prepared by SAN melanoma cells transfected with FKBP51/Flag. Cell lysates were immunoprecipitated with anti-Kat3B/p300 (IP p300) or anti-Flag (IP FKBP51). Immunoprecipitated and total lysates were then subjected to western blot with anti-FKBP51 or anti-p300. ( b ) ChIP performed with SAN melanoma cells, silenced (FKBP51 siRNA) or not (NS RNA) for FKBP51. An enrichment of DNA (region at −3450 from TSS of ABCG2 gene) can be observed in p300-immunoprecipitated chromatin (NS RNA) compared with IgG sample. Such an enrichment appeared to be reduced when FKBP51 was silenced (FKBP51 siRNA)
Article Snippet: Primary antibodies against FKBP51 (F-13; goat polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA), TGF- β (V; rabbit polyclonal; Santa Cruz Biotechnology), and Smad 2/3 (H465, rabbit polyclonal Santa Cruz Biotechnology) were used diluted 1 : 500; anti-FKBP51 (rabbit polyclonal; Novus Biologicals, Littleton, CO, USA), anti-FKBP51 (mouse polyclonal; Abnova, Taipei, Taiwan), anti-FKBP12 (N-19, goat polyclonal; Santa Cruz Biotechnology), anti-actin (I-19, goat polyclonal; Santa Cruz Biotechnology), anti-G3PDH (D16H11; rabbit monoclonal; Cell Signaling, Danvers, MA, USA), and anti-KAT3B/p300 (RW109; mouse monoclonal, Novus Biologicals) were used 1 : 1000.
Techniques: Transfection, Immunoprecipitation, Western Blot