Data_Sheet_1_Task-Specific Recognition Signals Are Located on the Legs in a Social Insect.docx

Task allocation ensures a high level of organization within social insect colonies. Workers reveal their task assignment through cuticular hydrocarbon (CHC) signals. The source and chemical composition of these signals are largely unknown. We ask whether task recognition signals are located on particular body parts of workers of Australian meat ants (Iridomyrmex purpureus). We analyzed the CHC profile on the antennae, legs, and abdomens of workers engaged in different tasks. Discriminant analysis showed that the leg profile is the best indicator of task identification. Behavioral assays confirmed this finding: workers typically reacted differently to non-nestmates engaged in different tasks, but not if the CHCs on the legs of their opponents were removed by a solvent. Lasso and Elastic-Net Regularized Generalized Linear Model (GLMNET) revealed which CHC components show the highest correlation in task and nestmate recognition, suggesting that social insects can simultaneously convey different CHC signals on different body parts, thereby allowing efficient signaling and signal perception.</p

Data_Sheet_2_Task-Specific Recognition Signals Are Located on the Legs in a Social Insect.xlsx

Task allocation ensures a high level of organization within social insect colonies. Workers reveal their task assignment through cuticular hydrocarbon (CHC) signals. The source and chemical composition of these signals are largely unknown. We ask whether task recognition signals are located on particular body parts of workers of Australian meat ants (Iridomyrmex purpureus). We analyzed the CHC profile on the antennae, legs, and abdomens of workers engaged in different tasks. Discriminant analysis showed that the leg profile is the best indicator of task identification. Behavioral assays confirmed this finding: workers typically reacted differently to non-nestmates engaged in different tasks, but not if the CHCs on the legs of their opponents were removed by a solvent. Lasso and Elastic-Net Regularized Generalized Linear Model (GLMNET) revealed which CHC components show the highest correlation in task and nestmate recognition, suggesting that social insects can simultaneously convey different CHC signals on different body parts, thereby allowing efficient signaling and signal perception.</p

Study timeline for bleomycin challenges, bioactive compound treatments, lung function assessments, and sampling time.

Study timeline for bleomycin challenges, bioactive compound treatments, lung function assessments, and sampling time.</p

Histopathology scoring data as assessed on histological H+E-stained sections sampled at post-mortem from the differentially treated lung-segments.

The differentially treated lung segments were the right medial (RM) lung-segments which were left untreated for healthy lung controls (Control), the right caudal (RC) and the left caudal (LC) lung-segments which were either infused with bleomycin without drug treatment (BLM) or infused with bleomycin and received 4 once-weekly doses of pinocembrin (BLM + PIN). The top panels show mean scoring data for ten sheep. The bottom panels show individual sheep data. Significance was determined using paired t-tests, *pMaterials and methods.</p

Representative Flash chromatography run of <i>Eucalyptus</i> extract showing the collected pinocembrin peak from 2.1 to 2.5 min.

The pinocembrin peak was collected from multiple runs and pooled to provide sufficient material for testing in the sheep trial. (PDF)</p

Quantitative analysis of the purity of pinocembrin isolated from <i>Eucalyptus</i> and used in the sheep trial.

A, HPLC-PDA data of pinocembrin purified from a Eucalyptus extract using Flash chromatography and injected at a concentration of 1 mg ml-1. B, HPLC-PDA data of an authentic standard of pinocembrin purchased from Sigma-Aldrich and also injected at a concentration of 1 mg ml-1. (PDF)</p

The effect of pinocembrin on lung parenchymal T cells.

(A) representative photomicrographs showing CD8+ and CD4+ T cells (arrows show examples of immuno-positive cells) in the lung parenchyma sampled from the differentially treated lung-segments at week 12. (B) graphs showing the average number of positive cells per field at 400x magnification for each differentially treated segment. The differentially treated lung segments were the right medial (RM) lung-segments which were left untreated for healthy lung controls (Control), the right caudal (RC) and the left caudal (LC) lung-segments which were either infused with bleomycin without drug treatment (BLM) or infused with bleomycin and received 4 once-weekly doses of pinocembrin (BLM + PIN). The left panels show mean lung segment data and the right panels show individual sheep data. Significance was determined using paired t-tests, **p<0.01, ***p<0.001, n = 10 sheep. Scale bars = 100 μm.</p

Lung function in the differentially treated lung segments as assessed at week 11 of the study.

The differentially treated lung segments were the right medial (RM) lung-segments which were left untreated for healthy lung controls (Control), the right caudal (RC) and the left caudal (LC) lung-segments which were either infused with bleomycin without drug treatment (BLM), or infused with bleomycin and received 4 once-weekly doses of pinocembrin (BLM + PIN). Part A shows mean data for Cseg (n = 10), which is a measure for how easy it is to inflate the lung segment. Part B shows individual sheep data. Part C shows percent change of Cseg at week 11 from baseline values taken at week 0 at the beginning of the study. Significance was determined using paired t-tests, *p<0.05, ***p<0.001, n = 10 sheep.</p

Neutrophils and inflammatory cells recovered from the bronchoalveolar lavage (BAL) fluid of the differentially treated lung-segments at week 12.

The differentially treated lung segments were the right medial (RM) lung-segments which were left untreated for healthy lung controls (Control), the right caudal (RC) and the left caudal (LC) lung-segments which were either infused with bleomycin without drug treatment (BLM) or infused with bleomycin and received 4 once-weekly doses of pinocembrin (BLM + PIN). The left panels show neutrophil data, and the right panels show inflammatory cell data, which included the sum of the percentages of neutrophils, lymphocytes, and eosinophils. The top panels show mean data for ten sheep. The bottom panel shows individual sheep data. Significance was determined using paired t-tests, *p<0.05, **p<0.01, ***p<0.001, n = 10 sheep.</p

S1 Data -

(XLSX)</p