XB-IMG-150672
Xenbase Image ID: 150672
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Fig. 7. Ascl1 is associated with VegT and mesendoderm genes. (A) Luciferase reporter assay shows that mutating the E-boxes in the proximal regulatory region of Xnr1 does not affect Ascl1 repression of the promoter activity induced by VegT. T1 and T2: functional T-boxes in the minimal Xnr1 promoter. E1 and E2: two putative E-boxes found in the Xnr1 minimal promoter by MEME algorithm. Crossed red rectangles indicate mutated E-boxes. s., significantly different (P<0.01). (B) Schematic of a minimal bix4 promoter with functionally characterized T-boxes and putative E-boxes highlighted. Green rectangles: T-boxes; red rectangles: putative E-boxes. 340 bp indicates the distance from the 3′ end of the E-boxes upstream of the 5′ end of the T-boxes. F, forward primer; R, reverse primer. (B′) ChIP-PCR results showing anti-MT-VegT ChIP recovers a bix4 promoter fragment from the T-box-containing region. (B′) ChIP-PCR results showing anti-MT-ASCL1 ChIP recovers a bix4 promoter fragment from the T-box-containing region. Values are means±s.d. (C) Western blots of anti-HA immunocomplexes detected by anti-Myc-tag or anti-HA antibodies indicating the association between ASCL1-δC and VegT. (D) A schematic depiction of MT-VegT serial deletions used in the CoIP/western blotting experiments shown in E,F. (E) HA-ASCL1-δC interacts with the T-box domain of VegT. (F) MT-ASCL1-NT interacts with Flag-VegT. (G) Results from CoIP followed by western blot analyses showing HDAC1 and VegT non-mutual exclusively associated with ASCL1. Image published in: Gao L et al. (2016) © 2016. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |