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VLDLR has a similar role with SR-B1 and LDLR in HCV entry.

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posted on 2016-05-06, 03:30 authored by Satomi Yamamoto, Takasuke Fukuhara, Chikako Ono, Kentaro Uemura, Yukako Kawachi, Mai Shiokawa, Hiroyuki Mori, Masami Wada, Ryoichi Shima, Toru Okamoto, Nobuhiko Hiraga, Ryosuke Suzuki, Kazuaki Chayama, Takaji Wakita, Yoshiharu Matsuura

(A) Relative mRNA expression of VLDLR in various tissues was determined using the NextBio Body Atlas application. (B) mRNA expression of SR-B1, LDLR, VLDLR and GAPDH in Huh7 cells and primary human hepatocyte (PHH) were determined by qRT-PCR. Relative expression levels of mRNA were calculated based on the expression level of GAPDH. (C) VLDLR-HA was exogenously expressed in parental, SR-KO, LD-KO and SR/LD-DKO Huh7 cells by infection with lentiviral vectors. Expressions of VLDLR in these cells were determined by immunoblotting analysis (upper panel). Parental, CD81 KO and SR/LD-DKO Huh7 cells expressing SR-B1, LDLR or VLDLR were infected with HCVcc at an MOI of 1 and intracellular HCV RNA levels were determined at 24 h post-infection (lower panel). (D) VLDLR-HA was exogenously expressed in CD81 KO, CLDN1 KO and OCLN KO Huh7 cells by infection with lentiviral vectors. Expressions of VLDLR, CD81, CLDN1 and OCLN in these cells were determined by immunoblotting analysis (upper panel). Cells were infected with HCVcc at an MOI of 1 and intracellular HCV RNA levels were determined at 24 h post-infection by qRT-PCR (lower panel). (E) SR-B1, LDLR and VLDLR were exogenously expressed in SR/LD-DKO Huh7 cells by infection with lentiviral vectors. Cells were infected with Con1-JFH1 or Jc1 at an MOI of 1, and intracellular HCV RNA levels were determined at 24 h post-infection by qRT-PCR. (F) Sera (100μl) from chimeric mice infected with HCV were inoculated into SR/LD-DKO Huh7 cells expressing either SR-B1, LDLR or VLDLR in 24 well plate. Intracellular HCV RNA levels were determined at 72 h post-infection. In all cases, asterisks indicate significant differences (*P<0.05; **P<0.01) versus the results for control cells.

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