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. Author manuscript; available in PMC: 2014 Dec 17.
Published in final edited form as: Cell Rep. 2014 Nov 26;9(5):1827–1840. doi: 10.1016/j.celrep.2014.10.063

Figure 5. Loss of Tet1 Expression is Associated with Decreased 5hmC and Increased 5mC Content in Kras-Transformed NIH3T3 Cells.

Figure 5

(A) mRNA levels were determined by RT-qPCR and normalized to NIH3T3 cells. Protein levels were determined by western blotting.

(B) Genomic 5mC and 5hmC levels were measured by DNA dot blot in the top panel. The blots were stained with methylene blue as a loading control in the bottom panel.

(C) Fas expression was determined by RT-qPCR and normalized to NIH3T3 cells.

(D) Methylation and hydroxymethylation levels of Fas promoter were analyzed by qPCR.

(E) Bisulfite sequencing for 5mC and TAB-seq for 5hmC. The percentages of 5mC or 5hmC were indicated.

(F) Tet1 chromatin occupancy was analyzed using Tet1 ChIP and qPCR. The data indicate that Kras transformation depresses Fas expression by converting the promoter from a 5hmC state to a 5mC state due to depletion of Tet1. All data are presented as mean ± SD. **, p < 0.01; ***, p < 0.001 in comparison to NIH3T3 cells. See also Figure S4.