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. 2002 Nov;76(22):11518–11529. doi: 10.1128/JVI.76.22.11518-11529.2002

FIG. 5.

FIG. 5.

Expression of the TGEV E protein in LLC-PK1 cell lines stably transformed with E protein expression plasmids. (A) Scheme of the E gene expression cassette in the pcDNA3.1(+) plasmid. CMV, CMV promoter; BGH pA, bovine growth hormone transcription stop signal and polyadenylation sequence; Neo, gene coding for neomycin resistance. (B) Detection of the mRNA for the E protein and β-actin by semiquantitative RT-PCR analysis in six cell lines (E-1, E-2, E-3, E-4, E-5, and E-6) stably transformed with the E gene. DNA amplification fragments were resolved by agarose gel electrophoresis and stained with ethidium bromide. (C) Western blot analysis of cell lysates from the six LLC-PK1 cell lines stably expressing the E protein, using an E-protein specific MAb. The position of the E protein band is indicated by an arrow. (D) Detection of E protein expression by immunofluorescence microscopy, using an E protein-specific MAb, in the six LLC-PK1 cell lines stably expressing the TGEV E protein. Mock, untransfected LLC-PK1 cells; TGEV, TGEV-infected LLC-PK1 cells.