Figure S5. The
bp292 mutation abolishes the methylation activity of EPG-11, Related to Figures 4,5,6 A-D. Loss of function of
prmt-5 causes no ectopic PGL granules in somatic cells (A,B) and does not enhance the defect in
epg-11 mutants (C,D) (compared to Figs. 1C,D). (A) and (C): DIC images of the embryos shown in (B) and (D). E-H. At postembryonic stages,
epg-11::gfp is expressed in pharyngeal cells (E,F, arrows) and vulval cells (G,H, arrow). (E) and (G): DIC images of the animals shown in (F) and (H). I. Life span of
epg-11 mutants is reduced compared to wild-type animals (log-rank test, p = 0.000). The median survival duration of wild type and
epg-11 mutants was 15 [95% confidence interval (CI) 13.099-16.901] and 12 (95% CI 11.389-12.611) days, respectively. The difference in survival was compared using the Kaplan-Meier method and log-rank tests. J. SEPA-1 fails to interact with EPG-11 in a GST pulldown assay. K-L. The interaction of EPG-11 with PGL-1 (K) or PGL-3 (L) is not affected by RNase A treatement in in vitro pulldown assays. The right panel shows the input protein levels used in the pull-down assay. Coom. Blue: Coomassie staining. M-N. Methylation levels of mutant PGL-3 RGG domains containing individual arginine-to-alanine mutations. The activity is shown as counts per minute (cpm) detected by a liquid scintillation counter. O-P. The anti-Rmeth antibody stains nuclei in early stage embryos. Q-R. The nuclear staining signal is largely absent in late stage embryos. S-T. The nuclear staining signal in early stage embryos is not affected in
pgl-1;
pgl-3. U-V. The nuclear staining is completely absent in
epg-11 mutants in early stage embryos. W-X. The nuclear staining signal in early stage embryos is not affected in
prmt-5 mutants. Y-Z. Germline P granules are stained by anti-Rmeth in
prmt-5 mutants. (O), (Q), (S), (U), (W) and (Y): DAPI images of the embryos shown in (P), (R), (T), (V), (X) and (Z), respectively. Bar: 10 μm (A-D, O-Z); 20 μm (E-H).