Research Paper Volume 13, Issue 22 pp 24580—24604

Figure 3. The BMPR2 mRNA levels were significantly reduced in the VHD cases compared with the healthy control samples. Potential diagnostic value was assessed for using the BMPR2 promoter DNA methylation levels or BMPR2 mRNA levels or both in the VHD plasma samples. (A) The levels of BMPR2 mRNA were quantified by qRT-PCR in the VHD cases (n = 36) and healthy control samples (n = 36). ^***P < 0.001. Since BMPR2 was not highly expressed, the difference in the Cq values (ΔCq) during qRT-PCR was used instead to compare its expression in the VHD patients with that of the controls (NC). (B) The BMPR2 mRNA levels were reduced in 97.2% (35/36) of the VHD cases compared with the healthy control samples. The BMPR2 mRNA levels were normalized to those of GAPDH in these samples. Horizontal axis, VHD patient number. Vertical axis, relative BMPR2 mRNA levels based on 2^−ΔΔCq after normalization. (C) The normalized BMPR2 mRNA levels in non-VHD healthy control (NC) samples. Horizontal axis, control sample number. Vertical axis, relative BMPR2 mRNA levels based on 2^−ΔΔCq after normalization. (D) Receiver operating characteristic curve (ROC) analysis was performed when the mean DNA methylation levels of 10 CpG sites at the BMPR2 promoter were used to distinguish plasma samples of VHD patients from those of normal controls. P < 0.001. AUC, area under curve. (E) ROC analysis of the BMPR2 mRNA levels was performed to distinguish plasma samples of VHD patients from those of normal controls. P < 0.001. (F) ROC analysis was performed when both mean BMPR2 promoter DNA methylation and mRNA levels were used in combination to distinguish plasma samples of VHD patients from those of the controls. 95% CI: 95% Confidence Interval. ^***P < 0.001.