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Fig. 3 | Virology Journal

Fig. 3

From: A substitution in the pre-S1 promoter region is associated with the viral regulation of hepatitis B virus

Fig. 3

A mutation in the pre-S1 promoter region Sp1 reduced transcription activity: (a-e) The levels of luciferase activity, mRNA expression, and protein expression of pre-S1 were evaluated using vectors containing the wild-type (WT), G2765A substitution (m5), and whole nucleotide mutation (m9) sequences. a pNL vectors inserted with the Sp1 promoter and a pGL4.54 TK promoter control vector were transfected into HepG2 cells. The levels of luciferase activity were normalized to the levels of the control vector. b Pre-S1 expression vectors were transfected into HepG2 cells. After 24 h, the total RNA was extracted, and polyA+ mRNA was purified and measured by qPCR. c-e Forty-eight hours after transfection with the pre-S1 expression vector, cell lysates were extracted and evaluated by immunoblotting using anti-HBs, anti-pre-S1 and anti-β-actin antibodies. The levels of L protein expression were normalized to the levels of S protein expression (d) or β-actin expression (e). f-k The wild-type (WT) or G2765A (m5) 1.3-fold HBV genome vector was transfected into HepG2 cells. The (f-h) cell lysates and (i-k) supernatants were evaluated by immunoblotting using anti-HBs, anti-pre-S1 and anti-β-actin antibodies. The levels of L protein expression were normalized to the levels of S protein expression (g, j) or β-actin expression (h, k). β-actin level of cell lysate was used for normalization of the supernatant sample isolated from the same well

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