## 2 Results  
### Bottom-Up Proteomics of Dinoponera quadriceps Venom  
We studied the protein content of _D. quadriceps_ venom using two proteomic strategies: (1) in-gel digestion and (2) in-solution digestion, followed by analysis with two different mass spectrometers. Initiating with the first strategy, as shown in Figure 1, 12 major clear protein bands were observed in the denaturing sodium dodecyl sulfate-polyacrylamideamide gel electrophoresis (SDS-PAGE). These bands were excised from the polyacrylamide gel, submitted to an in-gel digestion procedure, and analyzed by liquid chromatography coupled to a mass/mass spectrometer (LC-MS/MS). The output data were compared with the transcriptomic database of the _D. quadriceps_ venom gland. The list of venom proteins identified as toxins is presented in Table S1. Our gel-based proteomics analysis resulted in the unequivocal identification of nine major venom components that match perfectly with their respective transcripts (contigs) expressed in the _D. quadriceps_ venom gland (Table S1). After performing a homology search of databases, using the basic local alignment search tool (BLASTx), these main venom components were matched with the following polypeptides resolved by SDS-PAGE: gel band 1: venom dipeptidyl peptidase 4 (XP_014479089.1); gel bands 3 and 4: glucose dehydrogenase flavin adenine dinucleotide (FAD) (XP_014475855.1); gel band 5: protein yellow-like (XP_014473983.1); gel bands 6, 7, 8, 10, and 11: phospholipase A1 2-like(XP_014477282.1); gel band 7: hyaluronidase-like (XP_014477391.1); gel band 9: venom allergen 3-like (XP_014469499.1). In addition, we found three uncharacterized components: (i) gel bands 10, 11, and 12 corresponding to an uncharacterized protein LOC106750875 (XP_014486970.1); (ii) gel band 11 representing an uncharacterized protein LOC106751228 (XP_014487552.1); (iii) gel bands 11 and 12 representing an uncharacterized protein LOC105189919 (XP_011150625.1). Peptide fragments from these three uncharacterized proteins revealed a toxin with a Sol_i_2 (pheromone-binding protein/odorant-binding protein, OBP/PBP) domain (Table S1). Only in gel band 2, the giant ant venom-protein was not accurately identified. The complete list of venom polypeptides and their correspondent transcriptomic contigs that were identified through gel-based proteomics approach is shown, as aforementioned, in Table S1. The second strategy employed to study the _D. quadriceps_ venom was the solution-based proteomics approach, followed by analysis in two different mass spectrometers. This approach resulted in the identification of peptides corresponding to 61 venom-related transcript products in \"analysis #1\" (realized with the LTQ-Orbitrap Velos mass spectrometer) and 87 transcript products in \"analysis #2\" (realized with the Q Exactive Plus mass spectrometer) (Table S2). Both in-solution analyses led to the identification of 44 transcripts, of which seven were also identified in the in-gel analysis. These seven transcripts were venom dipeptidyl peptidase 4, glucose dehydrogenase FAD, protein yellow-like, phospholipase A1 2-like, hyaluronidase-like, venom allergen 3-like, and uncharacterized protein LOC106750875. These results reinforce the gel-based proteomics strategy, and also indicate the presence and expression of several protein components in the venom in quantities that is out of the dynamic range of Coomassieie brilliant blue (CBB)-staining of venom proteins separated by denaturing SDS-PAGE. For example, the results from these in-solution proteomic analyses showed the presence of less abundant polypeptides visible in the CBB-stained SDS-PAGE, such  
Figure 1: Resolution of _D. quadriceps_ venom by denaturing sodium dodecyl sulfate -polyacrylamide angle electrophoresis (SDS-PAGE). Dried venom (30 ug) was solubilized in SDS-PAGE sample buffer and separated in 12.5% T/2.6% C SDS-PAGE, under reducing conditions. The Coomassieie stained SDS-PAGE revealed the presence of 12 major, predominant bands in the crude venom.