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. Author manuscript; available in PMC: 2017 Oct 27.
Published in final edited form as: Oncogene. 2016 Oct 31;36(17):2435–2445. doi: 10.1038/onc.2016.399

Figure 3. HBV replication induces DNA demethylation of SALL4 in HepAD38 cell line.

Figure 3

(a) PCR quantification of SALL4 mRNA in HepAD38 cells grown without (−) HBV replication (D0) or with (+) HBV replication by tetracycline removal for 5, 10, 20 days (D5–D20). Results are from three independent RNA isolations performed in identical triplicates. Error bars denote S.D. *P< 0.05; **P< 0.01. (b) Immunoblot of SALL4 protein in HepAD38 cells −/+ HBV replication by tetracycline removal for D0–D20. (c) Bisulfite sequencing PCR results of SALL4 clones, using DNA from HepAD38 cells −/+ HBV replication by tetracycline removal for D0–D10. Open and closed circles denote unmethylated and methylated states, respectively. (d) QPCR of SALL4 mRNA following treatment with 10 µM 5-Aza-2-deoxycytidine (5AZA) or DMSO (vehicle control) for 48hr. Results are from three independent RNA isolations performed in identical triplicates. Error bars denote S.D. *P< 0.05; **P< 0.01.