Fig. 1.

TMEM176A expression and methylation status in human HCC cells. a Semi-quantitative RT-PCR shows TMEM176A expression levels in HCC cell lines. SNU182, SNU449, HBXF344, SMMC7721, Huh7, HepG2, LM3, PLC/PRF/5, BEL7405, SNU387, SNU475, and Huh1 are HCC cells. DAC: 5-Aza-2′-deoxycytidine; GAPDH: internal control; (-): absence of DAC; (+): presence of DAC. b MSP results of TMEM176A in HCC cell lines. U: unmethylated alleles; M: methylated alleles; IVD: in vitro methylated DNA, serves as methylation control; NL: normal peripheral lymphocytes DNA, serves as unmethylated control; H₂O: double distilled water. c BSSQ results of TMEM176A in LM3, SNU449, SNU387, and SNU475 cells. Double-headed arrow, MSP PCR product size was 159 bp and bisulfite sequencing focused on a 231-bp region of the CpG island (from − 388 to − 157) around the TMEM176A transcription start site. Filled circles: methylated CpG sites, open circles: unmethylated CpG sites. TSS: transcription start site