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Figure 1

From: Notch signaling facilitates hepatitis B virus covalently closed circular DNA transcription via cAMP response element-binding protein with E3 ubiquitin ligase-modulation

Figure 1

Notch inhibition markedly suppresses hepatis B virus (HBV) covalently closed circular DNA (cccDNA). (A) HBV Hirt DNA was extracted from HepG2.2.15.7 (i) and HepAD38 (ii) cells and detected by Southern blotting under the condition of no treatment, 85 °C for 5 min, or 85 °C for 5 min plus EcoRI 2 h digestion. (iii) EcoRI digestion of HepG2.2.15.7 cells for 2 h and 16 h after 85 °C treatment. (B) Jagged1 mRNA and protein levels were significantly decreased after transfection with Jagged1 shRNA, as detected by RTD-PCR (upper) and western blotting (below). (C) RTD-PCR quantification of cccDNA accumulation in HepG2.2.15.7 and HepAD38 cells transfected with lentiviruses encoding Jagged1 short hairpin RNA (shRNA). (D) Southern blot analysis confirming decreased cccDNA levels in HepG2.2.15.7 and HepAD38 cells transfected with Jagged1 shRNA. (E) No significant change in cell numbers at 0 and 120 h between 20 µM DMSO and DAPT treatment. (F) RTD-PCR quantification of cccDNA accumulation in HepG2.2.15.7 and HepAD38 cells administered 20 µM DAPT. (G) Southern blot analysis confirming decreased cccDNA levels in HepG2.2.15.7 and HepAD38 cells treated with 20 µM DAPT. cccDNA results are expressed as the number of cccDNA copies per genomic μg (mean ± standard deviation) from three independent experiments. *P < 0.05 and **P < 0.01 versus corresponding control shRNA values or 20 µM DMSO. Abbreviations: NT, no treatment; RC DNA, relaxed circular DNA; DSL DNA, double-stranded linear DNA; SS DNA, single-stranded DNA; shCON; Control-shRNA, shJAG1; Jagged1 shRNA; NS, not significant.

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