Figure 5: Methylation analysis of the TLR4 gene promoter in the wounds of DFU patients and controls was done. Bands of (A) control {C} samples and (B) DFU samples {P} were obtained after polymerase chain reaction of TLR4 (393 bp); (ND) indicates non-digested DNA samples, followed by digested DNA samples. (C) Bar graph showing methylation frequency of the AciI sites in TLR4 promoter in controls compared to DFU patients. Methylation pattern analysis of diabetic wounds revealed 74 % of them contained methylated CpG at –822 and –646 positions near a TATA box located close to the Sp-1 sites in the promoter of TLR4 compared to non diabetic control wounds containing only 50% methylated CpG.