Figure 3.

Analyses of vector genome copies (GCs) and meganuclease expression in consecutive macaque liver biopsy and necropsy samples collected at various time points after vector administration

(A) Vector GCs in the liver, as analyzed by qPCR of genomic DNA. (B) Expression of meganuclease in the liver, as measured by qPCR of total RNA isolated from liver samples followed by reverse transcription and presented as relative expression levels normalized by glyceraldehyde 3-phosphate dehydrogenase levels. RA2125-d284 was analyzed based on eight liver tissue samples collected at necropsy, two each from the left, right, middle, and caudate lobes. The means ± SEM are shown. Data from the first two biopsies of RA1866, RA1857, RA1829, RA2334, RA2125, and RA2343 have been previously published²⁴ and are included here for comparison purposes. (C) Images for ISH using transgene-specific probes (M1PCSK9) for liver biopsy samples at the indicated time points (ISH signal: red; nuclear DAPI stain: blue). Top row: images of third biopsies from animals previously described.²⁴ Middle row: images of sequential biopsies representative of the cohort that received steroid treatment. Bottom row: images of sequential biopsies representative of the AAV3B cohort. Scale bar, 100 μm.