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. 2015 Nov 12;30(3):1076–1086. doi: 10.1096/fj.15-280511

Figure 2.

Figure 2.

Southern blot showing episomal status of vectors. RKO mice were subretinally injected at postnatal day 3 (P3) with NP-cRho or NP-sgRho and retinas were collected at 1 mo PI. Total genomic DNA (15 µg) was digested with EcoRI, which has 2 restriction sties. A) Schematic of pEPI-MOP-cRho and pEPE-MOP-sgRho vectors with EcoRI restriction sites. Rho-ex1: rhodopsin exon1. B) The results showed distinct signals of either a 1.4 kb (cRho) or 5 kb (sgRho) rhodopsin fragment, which corresponds to the expected single band sizes, respectively. C) Membrane was striped and incubated with an S/MAR-specific probe. The results show a single distinct band of 5 kb of vector backbone, which matches with the size of the vector backbone. D, digested; U, undigested. D) Densitometric analysis of the Southern blot was used to assess mean copy number with S/MAR probe (standard curve was generated using the calculated copy numbers listed on the right).