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Figure 4 | Virology Journal

Figure 4

From: The E5 protein of the human papillomavirus type 16 down-regulates HLA-I surface expression in calnexin-expressing but not in calnexin-deficient cells

Figure 4

Calnexin interacts with the HPV16 E5 protein in cellular extracts. HEK-293T cells were transfected with AU1-tagged codon-optimised HPV16 E5, pCI-HPV16-E5-HA or corresponding empty vectors and lysed at 24 h posttransfection with CHAPS lysis buffer. A) Immunoblot showing the expression levels of the codon-optimised E5 gene (left panel) and of the viral E5 gene (right panel). Note the differences in the immunoreactivity signals despite the higher amount of total protein loaded in the non-optimised gene (100 μg vs 30 μg). B) Immunoprecipitations were performed using monoclonal anti-E5-tag (AU1) antibodies and proteins in the immune complexes were probed using anti-AU1 and anti-calnexin antibodies. C) Immunoprecipitations were performed using monoclonal anti-calnexin antibodies and proteins in the immune complexes were probed using anti-calnexin and anti-E5-tag (AU1) anti-bodies. D) Immunoprecipitations were performed using monoclonal anti-E5-tag (HA) antibodies and proteins in the immune complexes were probed using anti-HA and anti-calnexin antibodies. E) 2 Immunoprecipitations were performed using monoclonal anti-calnexin antibodies and proteins in the immune complexes were probed using anti-calnexin and anti-E5-tag (HA) antibodies. Molecular-mass markers in kDa are indicated at the left of the blots.

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Virology Journal

ISSN: 1743-422X