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. 2016 Oct 14;90(21):9632–9643. doi: 10.1128/JVI.01353-16

FIG 1.

FIG 1

Establishment of apoE knockout cell clone and permissiveness test. (A) Schematic representation of TALEN design and the sequencing of apoE knockout clone 2. A schematic diagram of the apoE gene sequence is shown (nucleotide positions are indicated). The locations of two TALEN arms are underlined, and the TALEN target is in bold. The sequence of apoE knockout clone 2 was aligned with that of the wild-type apoE gene (positive-strand sequence). (B) Western blot verification of apoE knockout in isolated cell clones derived from the Huh7.5.1 cell line. Proteins are specified on the right, and the molecular mass standards are marked on the left. Tubulin was used as a loading control. (C) Western blot verification of apoE3HA rescue cell line based on selected apoE knockout cell clone 2. (D) HCV replication efficiency in Huh7.5.1, apoE-KO, and apoE3HA cell lines was evaluated by electroporating JcR2a in vitro transcripts. (E) HCV production in Huh7.5.1, apoE-KO, and apoEHA cell lines was evaluated by electroporating Jc1 in vitro transcripts. Infectivity of Jc1 virus produced from Huh7.5.1, apoE-KO, and apoE3HA cells was detected by using a limiting-dilution assay in 96-well plates with Huh7.5.1 cells. (F) Detection of infectivity of Jc1 virus produced from Huh7.5.1, apoE-KO, and apoE3HA cell lines by using a limiting-dilution assay in 96-well plates. Shown are the means from 3 independent experiments; error bars indicate standard deviations from the means.