Figure 4

The specific expression of Venus at bone-like nodules in mouse primary osteoblast cultures infected with HDAd-hOC-Venus. (a) Mouse POBs were cultured for 6 days and then infected with either HDAd-hOC-Venus or HDAd-CAG-Venus at an MOI of 1,000. Two days later, the cells were stained with an anti-OC antibody. Negative control, uninfected mouse POBs. All images were taken under an IX81 microscope using a 20x objective lens. Ph: phase contrast images obtained with an exposure time of 10 ms in grayscale. Venus: GFP filter images obtained with an exposure time of 500 ms in green. Anti-OC: Immunostaining images of anti-OC detected by Alexa Fluor 594 with an exposure time of 1 s in red. DAPI: DAPI staining images with an exposure time of 1 s in blue. Merge: Merged images of the three colors. Scale bar: 50 μm. (b) FACS scatterplots of POBs infected with HDAd-hOC-Venus. The fractions with Venus-positive (+) and Venus-negative (−) cells were isolated by two-color FACS using hOC-Venus (Venus) and autofluorescence (PE-A) as the X- and Y-axes, respectively.