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. 2019 Aug 26;15(1-2):161–173. doi: 10.1080/15592294.2019.1656158

Figure 1.

Figure 1.

Reduced representation bisulphite sequencing data and DLGAP2 schematic demonstrating the region used for bisulphite pyrosequencing. (a) RRBS data for DLGAP2 from our prior study. This genome-scale analysis identified CpG sites that were differentially methylated in the sperm of cannabis users compared to controls. Results for nine CpG sites, intronically located in DLGAP2, that were significantly hypomethylated in the sperm of cannabis users compared to controls. (b) A gene schematic of DLGAP2. Exons are represented by the rectangular boxes, while the ovals represent CpG islands. The sequence identified by RRBS that was subsequently analysed here is identified in the inset. CpG site 7 (7*) was not initially identified in the RRBS data but included in further analyses, for a total of 10 CpG sites analysed. (c) Pyrosequencing assay validation (performed in triplicate) for DLGAP2. Data points and error bars represent the mean ± SEM, respectively. Error bars not shown had deviations too small for inclusion on the graph. (d) Pyrosequencing results showing the average level of DNA methylation for each of the ten CpG sites of DLGAP2 in human conceptal diploid testes tissues demonstrate that the intron 7 region of DLGAP2 is not an imprint control region. Shown are the mean ± SEM.* = p < 0.05; ** = p < 0.01; *** = p < 0.001.