Pitx3 mediates MTA1 and DJ1 interaction and recruitment to the TH promoter. (A) In vivo interaction of MTA1 and Pitx3 in SH-SY5Y cells by immunoprecipitation followed by Western blotting analysis. (B) Effect of Pitx3 knockdown on the ability of MTA1 or DJ1 to stimulate TH promoter Luc activity in SH-SY5Y cells. (C) Effect of DJ1 or MTA1 on the levels of TH mRNA in SH-SY5Y cells transfected with Pitx3 or control siRNAs. Values are the average of three independent measurements, and the SD from the mean is shown. (D) qPCR ChIP assay for recruitment of Pitx3 to the TH promoter region from −8,610 to −8,810. Values are the average of three independent measurements, and the SD from the mean is shown. (E and F) Effect of Pitx3 or control siRNAs on the recruitment of MTA1 or DJ1 on the TH promoter by real-time qPCR assays in SH-SY5Y cells. (G and H) Sequential double-ChIP followed by qPCR analysis performed with exponentially growing SH-SY5Y cells with Pitx3, MTA1, or DJ1 antibody. (I) Effect of MTA1 or DJ1 or both combined siRNAs on the recruitment of Pitx3 on TH promoter. (J and K) Effects of MTA1 or DJ1 and/or Pitx3 or CMV control along with full-length TH promoter-Luc activity in SH-SY5Y cells. (L) Effect of MTA1 or control siRNAs on the TH promoter-Luc activity by DJ1 or Pitx3 or together in SH-SY5Y cells. Values are the average of three independent measurements, and the SD from the mean is shown.