FIG. 1.

HBe was successfully transfected into U937 and Hmy2.CIR cells and expressed in these cells. (A) DNA extracted from Hmy2.CIR and transfected Hmy2.CIR cells were used as templates to amplify HBe DNA fragments. The result of DNA agarose gel electrophoresis showed that HBe DNA fragment was successfully integrated in Hmy2.CIR cells. Lane 1: empty lentivirus carrier—Hmy2.CIR; Lane 2: HBe-Hmy2.CIR; Lane 3: Hmy2.CIR; Lane 4 and Lane 5: negative control. (B) DNA extracted from U937 and transfected U937 cells were used as templates to amplify HBe DNA fragments. DNA agarose gel electrophoresis showed that HBe DNA fragment was successfully integrated in U937 cells also. Lane 1: empty lentivirus carrier—U937; Lane 2: HBe-U937; Lane 3: U937; Lane 4 and Lane 5: negative control. HBe DNA fragment is 552 bp in length. (C) HBe mRNA expression in Hmy2.CIR and U937 cells detected by reverse transcription PCR. Lane 1: empty lentivirus carrier—Hmy2.CIR; Lane 2: HBe-Hmy2.CIR; Lane 3: Hmy2.CIR; Lane 4 and Lane 5: negative control; Lane 6: DNA Marker; Lane 7: empty lentivirus carrier—U937; Lane 8: HBe-U937, Lane 9: U937; Lane 10 and Lane 11: negative control; Lane 12: DNA marker. The product of the reverse transcription PCR is 156 bp in length. This result proved that HBe DNA was successfully transcribed in transfected Hmy2.CIR and U937 cells. (D) Expression of HBe protein in transfected Hmy2.CIR cells detected by immunofluorescence. There was weak fluorescence in cytoplasm of HBe-Hmy2.CIR cells after incubation with HBe antibody and FITC-labeled second antibody (the first lane). It verified that HBe protein was weakly expressed in cytoplasm of HBe-Hmy2.CIR cells. Hmy2.CIR cells incubated with PBS was done as negative control. PCR, polymerase chain reaction. Color images available online at www.liebertpub.com/vim