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. 2006 Jan;80(2):810–820. doi: 10.1128/JVI.80.2.810-820.2006

FIG. 7.

FIG. 7.

Analysis of capsid conformations at the fivefold pores. Extracts from 293T cells transfected with an AAV2 wt or mutated genomic plasmid were fractionated by sucrose gradient centrifugation (10 to 30% sucrose). Aliquots of the 110S (DNA-containing particles) and 60S (empty or partially packaged particles) fractions were incubated at the designated temperatures for 5 min, applied to nitrocellulose membranes under nondenaturing conditions, and then reacted with antibodies A20, B1, A1, and A69. (A) Analyzed were the 110S fraction of lysates obtained from cells which were transfected with AAV2 wt genomic plasmids (pDM [VP Rep ssDNA]) as well as the 60S fractions obtained from lysates of cells transfected with a plasmid expressing AAV2 Rep and VPs (pDG [VP Rep]) or a plasmid expressing only VPs (pCMV-VP [VP]). (B) The 110S and 60S fractions of lysates obtained from cells which were transfected with AAV2 wt genomic plasmids (pTAV2.0 [wt]) were compared with the 60S fractions obtained from lysates of cells transfected with genomic plasmids of the packaging-deficient R432A and T329A/T330A mutants.