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. Author manuscript; available in PMC: 2014 Apr 1.
Published in final edited form as: Epigenomics. 2013 Apr;5(2):131–146. doi: 10.2217/epi.13.1

Figure 5. Functional analysis of CpG83 and CpG85 on Sox4 promoter activity.

Figure 5

Various Sox4 constructs in luciferase reporter vectors were transfected into murine embryonic maxillary mesenchymal cells. (A) Values in the y-axis represent normalized luciferase activities. The Sox4 reporter activities of all constructs were compared with that of the SV40 promoter (column #2; taken as 100%). Plasmids used for transfection included: (#1) pGL3 vector; (#2) SV40/pGL3; (#3) Sox4/pGL3; (#4) Sox4Δ/pGL3; (#5) Sox4Mut83/pGL3; and 6) Sox4Mut85/pGL3. Note the significant decrease in promoter activities of the last three constructs (#4–#6) when compared with that of Sox4/pGL3 (#3). (B) Indicates the salient features of the Sox4 plasmid constructs utilized. The location of the two differentially methylated regions and the CpG island is indicated in Sox4/pGL3, which harbors the 1.8-kb 5′ upstream region of Sox4. X denotes mutated CpG sites. Sox4/pCpGL was used for methylation analysis and harbors the 1.8-kb 5′ upstream region of Sox4 (only CpGs 83 and 85 are indicated in the figure). Transfections were repeated four times using duplicates and values are presented as mean ± standard deviation.

*p< 0.0001; **p < 0.0002.

DMR: Differentially methylated region; Mut: Mutation; SV: Simian virus 40.