PIK3R5 and metabolic dysfunction-associated steatotic liver disease: To confirm the results from WGBS, the methylation levels of CpG sites in promoter regions of PPARα, INS, CAMK2B, and PIK3R5, key genes in NAFLD, type II diabetes mellitus, insulin secretion, and cAMP signaling pathways, were determined using target next-generation bisulfite sequencing (tNGBS) (Figure 5).