Sites of transcriptional activity in the whole set of Drosophila melanogaster polytene chromosomes have been localized by means of fluorescent antibodies against DNA:RNA hybrid molecules and compared with results on 3H-uridine incorporation obtained earlier. The majority of large and small puffs with intensive 3H-uridine incorporation demonstrate bright fluorescence. Moreover, bright fluorescence is also observed for a large number of small puffs though the intensity of 3H-uridine incorporation is low. Some prominent puffs with high levels of 3H-uridine incorporation show weak fluorescence. Condensed bands, as a rule, do not show fluorescence. The regions that look like interbands under the light microscope are not real interbands, but consist of minibands visible only in the electron microscope (EM). However, a region that has been previously studied by EM and proven to be a real interband between two thick dark bands (100B3-100B4-5) showed fluorescence. These data support previous suggestions indicating a substantial contribution of transcriptional products from small puffs and interbands to the whole transcriptional system of polytene chromosomes.