Supplementary Information
Supplementary Figure Legends
Figure S1.
Related to Figure 2
B
.
Amino acid frequencies flanking predicted K
-
ε
-
GG for sites with
high and low SUMO
-
TR:control intensity ratios
in each replicate.
Figure S
2
.
Related to Figure 2
C
.
Relationship of diGly site intensity and reproducibility of
detection. Boxplots show the distributions of intensities for all diGly sites detected in each
replicate,
divided into categories depending on their detectability in other replicates.
Figure S
3
.
Related to Figure 2F. Localization of SUMOylation sites to IUPred2A
-
perdicteed
protein IDRs. Boxplots show the median IDR score for 10
-
aminoacid region flanking the central
diGly modified lysine for bona fide SUMO sites, compared to
randomly chosen lysine
in the same
protein and to background diGly sites.
Figure S
4
.
Related to Figure 4.
Confocal images of
ovarioles
from
control and SUMO knockdown
via MT
-
gal4 driven shRNA ovaries stained for Vasa
and DAPI. The arrowhead indicates collapsed
nuclei in mid
-
st
age egg chambers in SUMO knockdown ovaries.
Figure S
5
.
Related to Figure 5.
Western blot analysis (WB)
protein
SUMOylation in
Drosophila
S2 cells.
Indicated GFP
-
tagged proteins were overexpressed in S2 cells alongside 3xFlag3xHA
tagged SUMO, and cell lys
ates
were subjected to GFP immunoprecipitation followed by WB
detection first with anti
-
Flag antibody, and after stripping, anti
-
GFP antibody.
Asterisks indicate
the full length unmodified GFP
-
tagged protein form (red) or putative SUMO
-
modified form (green
).
Arrows indicate putative degradation products of unmodified (red) and SUMO
-
modified (green)
GFP
-
tagged protein. Images are representative from two independent biological replicates.
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chemistr
y
Acidic
Basic
Hydrophobic
Neutr
al
Polar
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SUMO-TR/control Intensity > 10
SUMO-TR/control Intensity <10
Replicate 1
Replicate 2
Replicate 3
Supplementary Figure 1
10
20
30
40
log2(Intensity_
exp#1+1)
10
20
30
40
log2(Intensity_
exp#2+1)
10
20
30
40
log2(Intensity_
exp#3 +1)
rep.1&2&3
rep.1&(2|3)
rep.1 only
rep.1&2&3
rep.2&(1|3)
rep.2 only
rep.1&2&3
rep.3&(1|2)
rep.3 only
Supplementary Figure 2
K~GG
random K
0.0
0.2
0.4
0.6
0.8
IUPred2A scores
median IDR score at diGly site+/-5 aa
K~GG
proteins with
SUMO site
proteins with
background GG site
Supplementary Figure 3
control
MTgal4>UASp-shSUMO
DAPI
vasa
merge
Supplementary Figure 4
100
150
250
75
50
*
*
*
*
250
75
20
50
37
25
100
150
anti-GFP
anti-Flag
Inputs
GFP-IP
GFP-Mael
GFP-Panx
GFP-Piwi
GFP-Spn-E
GFP-Mael
GFP-Panx
GFP-Piwi
GFP-Spn-E
<
Unsp.band
}
3xFlag3xHA-SUMO-
modified forms
*
*
*
*
*
Supplementary Figure 5