Figure 6.
Association of the dynein machinery with
oskar RNPs. (A–C) Localization of GFP tagged versions (green) of BicD (A), p50/dynamitin (B), and Dhc (C) with respect to oskar mRNA (magenta) in egg chambers during mid-oogenesis. Scale bar represents 10 μm. Note that the transient oskar accumulation in the center and at the anterior pole of early stage 9 oocytes is normal and disappears by the end of stage 9. Empty regions created upon rotating the images are shown in dark gray (A and B). (D and E) Quantification of oskar RNP association with the indicated proteins in the nurse cells and the oocyte as a function of RNA copy number. The number of analyzed oskar RNPs was 12,945 (GFP-Dhc, nurse cells), 34,428 (GFP-p50/dynamitin, nurse cells), 8,573 (BicD-GFP, nurse cells), 27,936 (GFP-Dhc, oocytes), 37,992 (GFP-p50/dynamitin, oocytes), and 20,456 (BicD-GFP, oocytes). (F–G′) Quantification of oskar RNP association with GFP-Dhc and BicD-GFP in oocytes of indicated genotypes. 29,428, 32,132 and 33,638 oskar RNPs were analyzed in control, stau RNAi and Egl overexpressing oocytes, respectively (F′). 20,456 and 32,517 oskar RNPs were analyzed in control and stau RNAi oocytes, respectively (G′). Error bars represent 95% confidence intervals, and the size of the circles is proportional to the relative abundance of each category of oskar RNP within the overall population. In D, F, and F′, datapoints are slightly offset in the x-axis to facilitate comparison. Triangles indicate that the fraction of GFP-positive oskar RNPs is not significantly different from zero (P > 0.01, one sample t test, D–G).