Figure 5. Expression characteristics of
dual promoter vectors constructed using rhodopsin kinase (RK) and
interphotorecepter binding protein (IRBP)156 promoters.
pFIN-RK-GFP-IRBP156-CHER-WPRE (3.3×107 vector genomes/µl; A-E
and F-J), pFIN-IRBP156-CHER-RK-GFP-WPRE (1.5×109
vector genomes/µl; K-O and P-T),
pFIN-RK-GFP-RK-CHER-WPRE (1.85×107 vector genomes/µl; U-Y
and Z-DD), or pFIN-IRBP156-CHER-IRBP156-GFP-WPRE (1.7×109
vector genomes/µl; EE-II and JJ-NN) lentivirus was
injected into the developing neural tubes of E2 chicken embryos in ovo.
A minimum of four retinal whole mounts were examined for each virus.
Retinal regions shown in the figure were selected to illustrate the
range of transgene expression characteristics observed in infected
cells. Each region was photographed twice using the exposure duration
shown in the lower left of each panel and filters appropriate for
detection of CHER or GFP. Each row in the figure shows one selected
region. The merged images (C, H, M, R, W,
BB, GG, LL) were analyzed using the
co-localization module of the Zeiss AxioVision Image Suite. The results
of these analyses are expressed as the percent of the transduced area
in the image (pixels) containing co-localized GFP and CHER (yellow bar)
or GFP (green bar) or CHER (red bar) fluorescence alone. (D, I,
N, S, X, CC, HH, MM). The
images shown in E, J, O, T, Y, DD,
II, NN were extracted from the merged images (C-LL)
show only those areas of the merged image in which GFP was
co-localized with CHER. The scale bar shown in A is applicable
to all images and equals 50 µm.