Secondhand smoke in combination with ambient air pollution exposure is associated with increasedx CpG methylation and decreased expression of IFN-γ in T effector cells and Foxp3 in T regulatory cells in children

Table 4 Oligonucleotides used for bisulphite-specific polymerase chain reaction and pyrosequencing of IFN-γ

CpG Site*	Details	Sequences
−295	Forward	5′-[Biotin]TTTGTAAAGGTTTGAGAGGTTTTAGAAT-3′
	Reverse	5′-CAAACCCATTATACCCACCTATACCA-3′
	Sequencer	5′-TTTTATACCTCCCCACTT-3′
−186	Forward	5′-TTAGAATGGTATAGGTGGGTATAATGG-3′
	Reverse	5′-[Biotin] TATTATAATTAAAATTTCCTTTAAACTCCT-3′
	Sequencer	5′-GGGTATAATGGGTTTGTT-3′
−54	Forward	5′-GGGTTTGTTTTATAGTTAAAGGATTTAAGG-3′
	Reverse	5′-[Biotin] AATCAAAACAATATACTACACCTCCTCTAA-3′
	Sequencer	5′-TATTTTATTTTAAAAAATTTGTG-3′
+122~	Forward	5′-[Biotin] TTTTGGATTTGATTAGTTTGATATAAGAA-3′
+128	Reverse	5′-AAAACCCAAAACCATACAAAACTAAAA-3′
	Sequencer	5′-CTAAAAAACCAAAATATAACTTAT-3′
+171	Forward	5′-[Biotin] TTTTGGATTTGATTAGTTTGATATAAGAA-3′
	Reverse	5′-CATTTTCAACCACAAACAAATACTATTAA-3′
	Sequencer	5′-ACAACCAAAAAAACCC-3′

*CpG sites indicate nucleotide position in relation to transcription start. Symbols – and + indicate upstream and downstream of transcription start, respectively. CpG, cytosine nucleotide neighboring guanine.

ISSN: 1868-7083