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. 2023 Jul 12;14:1213285. doi: 10.3389/fimmu.2023.1213285

Figure 1.

Figure 1

(A) Illustration of the fusion protein containing an IgE signal peptide (SP), a modified E6 (non-oncogenic version) linked to a modified E7 (non-oncogenic version) via a Furin cleavage site. (B) Representative Western Blot assay detecting the HPV16 E6-E7 fusion protein (~31 kDa) with either an Anti-E6 or an Anti-E7 antibody on lysates from HEK293T cells transfected with 0.5 μg mRNA, and analyzed at 6, 24, 48, 72 hours, 6 and 8 days post-transfection. Quantification of band intensity at the indicated time points is shown on the right. (C) Representative Western Blot assay detecting the HPV16 E6-E7 fusion protein (~31 kDa) with either an Anti-E6 or an Anti-E7 antibody on lysates from HEK293T cells transfected with different amounts of mRNA (0, 0.5, 10, 25, 50, 150 and 500 ng/well), and analyzed at 24 hours post-transfection. Quantification of band intensity at the indicated time points is shown on the right. (D) Western Blot assay detecting the non-glycosylated HPV16 E6-E7 fusion protein (~31 kDa) in lysates of mRNA-transfected HeLa cells using Anti-HPV E7 antibody. The non-glycosylated HPV16 E6-E7 fusion protein was obtained by the inhibition of in-situ glycosylation with different concentration of Tunicamycin (μg/mL) or by protein de-glycosylation with PNGase (500U/μl) treatment of the cell lysates. Transfection controls: (+) untreated lysate from mRNA-transfected cells;(-) lysate from non-transfected cells.