Localization of 5hmC peaks in the INSIG1 gene promoter by analysis of 5hmC sequencing, confirmation of the 5hmC peaks by hMeDIP and MedIP analysis, and knockdown of INSIG1 mitigated the process of hypoxia-induced epithelial-mesenchymal transition. (a) Visualization of 5hmC signal in the INSIG1 promoter. The y axis is normalized read counts. The regions analyzed for 5hmC and 5mC levels (b,c) were labeled as dark brackets. (b) Confirmation of increased 5hmC peaks in the promoter region of INSIG1 gene using hMeDIP analysis. N; normoxia; H: hypoxia. (c) Confirmation of decreased 5mC peak in the promoter of INSIG1 gene using MeDIP analysis. The asterisk (*) indicates statistical significance (P <0.05) between experimental and control clones. The FADU scrambled control clone under normoxia was chosen as the control condition in (b) and (c). Error bars indicate standard deviations (s.d.) of duplicate analysis by PCR (b,c). (d) Knockdown of INSIG1 mitigated hypoxia-induced epithelial-mesenchymal transition.