Figure 3.
The kinase inhibitor 6-DMAP causes an additional period of BrdU incorporation in 43B-UAS–Cyclin E larval salivary glands. Salivary glands were dissected from second instar 43B-UAS–Cyclin E larvae and incubated for 30 min in tissue culture medium with or without 10 mM 6-DMAP. The glands were then washed six times for 5 min with medium free of 6-DMAP and labeled for 30 min in culture medium with BrdU. (a) Untreated glands show only some small dots of BrdU incorporation (arrows). (b) Treatment with 6-DMAP resulted in BrdU incorporation in both dots (arrows) and entire nuclei (arrowheads). BrdU was detected using rhodamine-conjugated secondary antibodies and the images captured using a CCD (charge-coupled device) camera.