Xenbase Image ID: 158332

Figure 3 The 11-bp NRE Binds to β-Catenin and Tcf (A) We looked for the factor(s) that bind to the 11-bp NRE. (B) We performed EMSA using Xenopus egg extract and a 30-bp probe containing the 11-bp NRE. IR denotes infrared dye used to tag the DNA probe. (C–F) EMSA analysis. Every panel shown comes from a single gel. Each experiment was repeated two to four times. In all gels, the red arrow indicates the specific EMSA band. (C) Left gel: competition with excess, unlabeled wild-type probe (lanes 3 and 4) and excess, unlabeled probe containing the m11 mutation (lanes 5 and 6). Right gel: EMSA using a different batch of Xenopus extracts and resolved using a lower-percentage gel. (D) Competition with excess, unlabeled wild-type probe (lanes 3 and 4), WRE probe (lanes 5 and 6), and mutant WRE probe (lanes 7 and 8). See also Figure S2A. (E) Competition with polyclonal XTcf3 antibody against the C-terminal of XTcf3 (XTcf3c, lanes 2 and 3) and against the N-terminal of XTcf3 (XTcf3n, lanes 4 and 5). (F) Competition with polyclonal antibody against β-catenin. Red arrow, the specific EMSA band; green arrows, a supershift and a smear downshift. See also Figure S2B. (G) Chromatin immunoprecipitation using β-catenin antibody. Genomic DNA was isolated from stage 10 Xenopus embryos, sonicated, and pulled down with β-catenin antibody. Lane 1: PCR amplification from the 11-bp NRE region. Lanes 2–4: PCR amplification from regions containing WRE in the siamois promoter. Image published in: Kim K et al. (2017) Copyright © 2017. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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