a. Fractionation of isopycnic Nycodenz density gradients of post-nuclear yw embryo lysate. Protein and Nycodenz concentration for every fraction is plotted.
b. Western blot detection of indicated proteins in gradient fractions. A short and a long exposure (long exp.) for Aub is shown. Uncropped gels for panels b, d and e can be found in Supplementary Figure 1.
c. Heat map of levels of indicated germ plasm mRNA determined by qRT-PCR, normalized to spiked luciferase RNA, and with fraction 2 as a reference.
d. Western blot detection of Aub in indicated diluted Nycodenz fractions used for Aub RNA IP.
e. Electrophoretic analysis on denaturing polyacrylamide gels of 32P-labeled small RNAs immunoprecipitated with Aub from indicated gradient fractions. A bracket denotes piRNAs, detected primarily in fractions 6 and 7 (asterisk: 2S rRNA).
f. Bar plot showing -fold enrichment (over fraction-extracted total RNA) of indicated germ plasm mRNAs in Aub IPs from gradient fractions, measured by qRT-PCR. Luciferase mRNA was used as a spike.