Figure 6

YFP is expressed in the epithelial cells of the terminal airways. YFP expression patterns observed using confocal microscopy of mouse lung parenchyma cryosections 24 hours after (a, b and c) dosage of 75 μg with post-nebulized pVR1020–YFP in 0.9% NaCl aqueous solution showing (a) YFP expression in a cluster of epithelial cells where lung cell nuclei stained with DAPI dilactate appear blue while cells expressing YFP appear green (scale bar =5 μm); (b) another view of a part of the YFP-expressing cell region shows the YFP distribution in the cells with appropriate filtering to (b) show only the nuclei and (c) YFP (scale bar =5 μm). (d) Western blot of the supernatant obtained from homogenized mice lungs harvested (left lane) 24 hours post-transfection with 300 μg pVR1020–YFP plasmid that was SAW nebulized at 30 MHz and instilled, compared to (right lane) the supernatant from untreated mice lungs. The YFP protein appears clearly at 27 kDa. The additional protein bands present at 50 and 37 kDa for both groups are due to the polyclonal nature of the rabbit anti-YFP antisera used as the probe where cross-reactivity was induced across other serum proteins.