Figure S1. Gas-chromatogram showing the typical chemical profile of a Pachycondyla inversa worker.

Figure S2. Temporal analysis of task profiles and nest cavity locations. Distributions of r^2-values
(proportions of variation explained by matrilines within each colony) in the resampled data sets (the histogram bars), and data sets consisting of 10 consecutive observations (hatched vertical lines). 96% of the temporally structured observations are within the 95% CI of the random distribution, and all of the within 99% CI, which suggests absence of temporal structure in the data.
Task profiles
Colony 1 Colony 2
0.1
0.2
0.3
0.4
0.5
0.6
0.7
Colony 3
0.0
0.1
0.2
0.3
0.4
0.5
Colony 4
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.0
0.1
0.2
0.3
0.4
Nest cavity locations
Colony 1
Colony 2
0.1
0.2
0.3
0.4
0.5
0.6
Colony 3
0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45
Colony 4
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.05
0.10
0.15
0.20
Proportion of variance explained (r^2) by matrilines within colonies
0.25

Table S1. Numbers of alleles (nall) and expected heterozygosities (He) for each locus and each colony.
Locus Pin6-1 Pin2 Pin6-2 Pin14 Pin61
Colony 1
Colony 2
Colony 3
Colony 4
Colony 5
nall 9
He 0.786
nall 10
He
nall 9
He 0.842
nall 7
He
nall 6
He
0.864
0.813
0.665
5
0.662
7
0.580
6
0.701
4
0.685
4
0.652
8
0.784
14
0.846
10
0.866
7
0.830
4
0.424
9
0.827
9
0.850
8
0.826
7
0.786
5
0.647
6
0.690
12
0.818
Not genotyped
Not genotyped
6
0.809
5
0.653

Table S2. Matriline shares presented as numbers of workers belonging to each matriline in the experimental colonies, the effective number of queens (NQe) and relatedness among workers (rw) in the colony. NQe calculated as 1/
 qi^2, where qi is the proportion of offspring produced by the ith queen, assuming single mating. rw is calculated as, 3 /4NQe, assuming unrelated queens. Some individuals died between chemical samples and observations. Specifically, in colony 2, some individuals of matriline Q4 died after the chemical samples were taken, and the number of individuals with behavioural observations in that matriline dropped below three. This means that the number of matrilines analysed for colony 2 is different in the chemical and behavioural data sets.
2
3
Colony
1
4
5
Q1
11
15
9
20
32
Q2
6
9
8
16
Q3
6
7
6
12
Q4
3
5
4
Q5
2
1
1
Q6
2
U
4
NQe
5.1
3.6
4.0
2.9
1 rq rw
0.15
0.21
0.19
0.26
0.75

Table S3. Compounds identified and their loadings on the PC axes used in the analyses. For the PC-axes, percentages of variation explained and eigenvalues
(

) are given. Cut-off of
 was used for retaining PC axes in further analyses.
15
16
17
18
19
20
21
22
23
8
9
10
11
12
13
14
2
3
4
5
6
7
Peak Compound number name
1 Unknown
11-MeC
25
3-MeC
25
10+12-MeC
26
6-MeC
26
4-MeC
26 x-MeC
27:1
C
27
0.04
9+11+13-MeC
27
-0.17
7-MeC
27
5-MeC
27
0.28
0.05
9,y-diMeC
27
3-MeC
27
C28
-0.20
-0.01
0.29
PC1 PC2 PC3 PC4 PC5
(18%,

= 6.2) (10%,

= 1.7) (8%,

= 1.2) (8%,

= 1.2) (7%,

= 0.9)
-0.22 0.04 -0.07 -0.36 0.08
-0.45
-0.17
-0.16
-0.41
-0.18
-0.10
0.12
0.13
-0.11
-0.03
-0.16
0.27
-0.03
0.18
-0.30
-0.05
-0.21
0.49
0.10
-0.04
-0.27
0.13
0.05
-0.32
0.05
0.07
-0.03
0.01
-0.29
0.06
-0.18
-0.10
0.06
-0.27
0.06
0.04
-0.08
-0.27
-0.01
0.37
0.29
0.20
-0.08
-0.17
-0.21
0.08
-0.03
0.19
0.10
0.00
-0.25
0.02
0.01
-0.11
-0.10
-0.08
-0.09
-0.08
3,11-diMeC
27
0.09
10+12-MeC
28
0.11
4-MeC
28
2-MeC
28
Unknown
C29
11+13-MeC
3,11-diMeC
29
Unknown
29
0.17
0.15
0.19
0.25
0.13
0.23
0.09
-0.27
0.03
-0.18
0.00
-0.26
-0.06
0.09
0.71
0.16
-0.11
0.06
0.06
0.17
-0.08
-0.05
0.11
-0.38
-0.10
0.32
0.06
-0.08
0.22
0.01
-0.22
-0.07
0.07
0.54
-0.13
0.02
0.26
-0.57
0.23
0.06
0.26
-0.19
0.53

Table S4. Percentages of occasions when queens and brood were observed in each nest cavity. For each queen, these do not add to 100% since a queen may have been observed outside a nest cavity. For brood, they add up to >100% when brood has been split into several cavities on some occasions. The bottom row “Separated” gives the proportion of times when a queen was observed in a cavity where no brood at all was present, and for brood, the proportion of occasions when at least some brood was in cavity where no queens were present. Numbers of brood items in different cavities where not recorded, so we cannot tell whether the brood that was occasionally separated from queens represented a large proportion of brood.
3
4
1
2
1
2
Colony 1
Cavity
3
4
Separated 1
55
6
Colony 2
Cavity Q1
55
4
1
Q2
Q1 Q2
39 39
3
74
14
Separated 13
3
87
4
16
Colony 3
Cavity
3
4
1
2
Q1 Q2
3 1
16
71
6
16
74
4
Separated 4 5
Q3
3
38
55
3
2
Q3
9
77
7
14
Q3
0
14
72
9
3
Q4
Q4
1
93
3
3
Q4
0
7
74
12
1
Q5
Q5
94
3
12
Q5
0
17
72
10
2
Q6
Q6
100
0
Q6
Q7
Q7
6
77
12
1
Q7
Brood
4
43
67
23
19
Brood
12
10
100
10
25
Brood
3
33
90
23
16

Colony 4
Cavity
3
4
1
2
Separated 1
1
87
10
Q1 Q2
94
4
5
Colony 5
Cavity
3
4
1
2
Separated 2
Q1 Q2
61
39
48
46
6
Q3
1
91
7
3
Q3
Q4 Q5 Q6 Q7
Q4 Q5 Q6 Q7
Brood
4
97
22
11
Brood
1
70
46
9