Fig. S4

Fig. S4

Sorting of Intestine-Specific Cells and Requirement of Demethylase for Aberrant Expression of aldh1a2, fabp2, pcna, cyclind1 and pitx2 in apc Mutant Zebrafish Embryos, Related to Figure 4

(A) FACS sorting parameters for GFP negative apc^wt embryos, gutGFP;apc^mcr and gutGFP;apc^wt embryos.

(B) Fabp2 expression in GFP positive and negative sorted cells from gutGFP;apc^mcr and gutGFP;apc^wt embryos.

(C) RT-PCR showing aldh1a2 and fabp2 expression in apc^mcr and apc^wt embryos injected with control Mo/ aaa Mo/ mbd4+tdg Mo. Error bars are +/- SD.

(D) Quantitative RT-PCR for AID, MBD4 and GADD45A in ALDH positive or negative fractions (FACS sorted using aldefluor assay) of DLD1 and HT29 cells. Fold enrichment was assessed over 18S expression levels and values from ALDH negative fraction were normalized as 1.

(E) Relative expression of cyclind1, aldh1a2 and fabp2 in the intestines of adult zebrafish treated with DMSO or DEAB for 7 days.

(F) Whole mount in situ staining for pcna, cyclind1 and pitx2 in apc^mcr and apc^wt zebrafish embryos injected with control Mo, aaa Mo, mbd4/tdg Mo, pitx2 Mo, or dnmt1 expression plasmid.

(G and H) Whole mount in situ for krox20 and hoxb4 (panel G, krox20 in red and hoxb4 in blue at 24hpf) and dlx2 and sox2 (panel H, dlx2 at 24hpf and sox2 at 72hpf) in apc^mcr and apc^wt embryos.