Figure 1 | Scientific Reports

Figure 1

From: Vector uncoating limits adeno-associated viral vector-mediated transduction of human dendritic cells and vector immunogenicity

Figure 1

Early steps in iDC transduction by AAV vectors. (A) Entry efficiency of AAV vectors derived from different serotypes. iDC were incubated with AAV vectors encoding for enhanced green fluorescent protein (GFP) in single-stranded vector genome conformation at a GOI of 104 for 48 hrs. Cells were harvested by extensive trypsin treatment to remove membrane-bound particles22. Total DNA was extracted and analyzed by qPCR using primers specific for GFP (vector genome) and for the host cell gene (β-globin), respectively. Results are presented as the mean GFP-to-β-globin ratio (R.U.). Error bars represent standard deviation (n = 3). (B) AAV2 vectors are delivered to the nucleus. iDC were incubated with AAV2 vectors at a GOI of 104 for 7 hrs. Cells were fixed, permeabilized, and incubated with antibodies directed against the AAV capsid (A20) and nuclear Lamin B1, respectively. The nucleus was stained with DAPI. Cells were imaged by confocal microscopy. Arrow heads indicate A20 signals in the nucleus.

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