Fig. 3
Production of additional recombinant proteins using FlexiBAC. a Human Bone morphogenetic protein 4 (BMP4) was expressed in insect cells using the two different FlexiBAC viral backbones that differ in their expression of recombinant furin: DefBac and DefBacFur+. BMP4 levels in the conditioned media were analyzed by western blot using an anti-BMP4 antibody. b Human Growth/differentiation factor 15 (GDF15) was expressed in insect cells using DefBac and DefBacFur+. GDF15 levels in the conditioned media were analyzed by western blot using an anti-GDF-15 antibody. c The ecto-domain of canine E-cadherin was expressed in insect cells using DefBac and DefBacFur+. E-cadherin levels in the conditioned media were analyzed by western blot using an anti-E-cadherin antibody. d Coomassie-stained gels depicting 6xHis-tagged SPD-5::GFP expressed with the Bac-to-Bac system (ThermoFisher Scientific) or with DefBac. In both lanes, SPD-5::GFP was purified from insect cell lysate using Ni-NTA affinity chromatography, then eluted with 250 mM imidazole. e All proteins in (B-E) were expressed in SF9 insect cells using the FlexiBAC system. Coomassie-stained gels depicting purified C. elegans centrosome proteins. For more information, see [17, 27]. f Coomassie-stained gels of purified human proteins that contain long stretches of intrinsically-disordered regions. For more information, see [22, 25]. g Coomassie-stained gels depicting purified yeast prion-like proteins. MC and C represent middle and C-terminal domains of Sup35. For more information, see [24]. h Coomassie-stained gel of purified PGL-3, a protein that constitutes P granules in C. elegans embryos. For more information, see [23]