Figure 9

Schematic diagram and sequence analysis of open reading frame of human IFNAR1 by RT-PCR from the S-5/15 and R-15/1, R-17/1 and R-24/1 cells. (A) Location of sense and antisense primers used for RT-PCR analysis of IFNAR1 coding sequence. The human IFNAR1 protein is 557 amino acids long that consist of an N-terminal extracellular cytokine-binding (EC) domain (436 aa), a hydrophobic transmembrane (TM) domain (21 aa), and C-terminal intracytoplasmic (IC) domain (100 aa). F1 and F2 are the two PCR fragments amplified for final overlapping PCR of full-length IFNAR1. (B) The amino acid sequence of IFNAR1 of S-5/15 cells perfectly matches with the reference wild type sequence of IFNAR1 m RNA (Accession no- NM_000629). The R-15 and R-24 series resistant Huh-7 clone show an N-terminal deletion of 58 amino acids starting from 68 aa to 125 aa in the IFNAR1. The R-17 series resistant Huh-7 cell clones show deletion of 50 amino acids staring from 382aa to 431 aa in the SD4 domain of IFNAR1. The dotted line indicates the deletion portion in the sequence.