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. 2020 Nov 5;17(11):e1003229. doi: 10.1371/journal.pmed.1003229

Fig 4. Validation results of the array using bisulphite pyrosequencing.

Fig 4

(A–D) Validation of cg16536918, in the AVP gene, by pyrosequencing, showing (A) the location of the CpG in the AVP DMR found associated with GDM, (B) the significant association with 1-h PG levels, (C) GDM status, and (D) the correlation between methylation levels measured on the pyrosequencer and beta values on the array. (E–H) Validation of cg26953232, a CpG in the DMR, found associated with 1-h PG levels, showing (E) the location of the CpG in the genome, (F) the significant association with 1-h PG, (G) GDM status, and (H) the correlation between methylation levels measured on the pyrosequencer and beta values on the array. (A+E) Diagrams of the dmCpGs and the location of the (A) AVP and (E) BRD2 pyrosequencer assay. Red circles indicate CpGs on the array identified as part of the DMR. Purple circles indicate extra CpGs measured on the pyrosequencer, with the location of the 3 sequencing primers used (Seq1–3). AVP, arginine vasopressin; BRD2, Bromodomain Containing 2; CpG, cytosine-phosphate-guanine; dmCpG, differentially methylated CpG; DMR, differentially methylated region; GDM, gestational diabetes mellitus; PCR, polymerase chain reaction; PG, plasma glucose.