Extended Data Figure 5. Partial purification of Aub RNPs from early embryo supports piRNA independent binding of germ plasm mRNAs by Aub.
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.