Aquaporin 5 Expression in Mouse Mammary Gland Cells Is Not Driven by Promoter Methylation

Figure 3

Analysis of AQP5 promoter methylation. (a) DNA methylation of 49 CpG sites found in the 289 base pair region located ~70 bases from the start codon of the AQP5 promoter, measured by bisulfite sequencing of individual clones (total methylation percentage shown in black). A single CpG site (CpG#24) was measured in tens of thousands of molecules with bead-emulsion amplification (methylation percentage shown in green). Nondividing cells without Dex and with Dex treatments for 2 h and 4 h were analyzed. (b) Changes in AQP5 mRNA expression analyzed with one-step qPCR. Total mRNA was extracted from Matrigel-treated cells without Dex and with Dex treatments for 2 h and 4 h. AQP5 expression is normalized to GAPDH and shown relative to no Dex treatment. Bars represent averages and confidence intervals; ; alpha = 0.05. Significance was tested between control (−Dex) and differently treated samples. (c) Summary of AQP5 mRNA expression and promoter methylation.

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