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. 2021 Jan 17;30(3-4):245–254. doi: 10.1038/s41434-020-00218-6

Fig. 1. Generation of a dual hybrid AAV vector expressing VWF in HUVEC cells.

Fig. 1

a Schematic representation of the dual hybrid AAV vector. ITR inverted terminal repeats for AAV packaging, ICAM2 intercellular adhesion molecule 2 promoter, 5′ VWF 5′ region of the VWF coding sequence, from nucleotide +1 to +4059, SD splicing donor site, AK recombinogenic region from F1 phage genome, SA splicing acceptor site, 3′ VWF 3′ region of the VWF coding sequence, from nucleotide +4060 to +8442. b On the top, scheme of the primers used to amplify VWF coding sequence from vector genome. On the bottom left, PCR analysis of the VWF amplicons. On the bottom right, a schematic representation of the amplified viral genomic region is depicted. c Representative western blot analyses on HUVEC cell lysates and conditioned media samples collected 72 h post transduction. On the bottom, histogram reporting the western blot quantification via densitometric analysis. Cntrl, untransduced HUVEC cells. Data are reported as mean ± SD from experiments performed in triplicate, **p < 0.01 (one-way ANOVA). d Representative western blot analyses on Huh7 and HEK293 cell lysates and conditioned media samples collected 72 h post transduction. Cntrl, untransduced cells.