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. 2019 Feb 10;1:100003. doi: 10.1016/j.yjsbx.2019.100003

Fig. 4.

Fig. 4

Fig. 4

Examples of multiprotein complexes expressed using the SmartBac system. (a) Fluorescence signals for tagRFP (top) and EGFP (bottom) detected from Sf9 cells transfected with BVE1S547 and BV2863 (see Table 1). (b) Coomassie-stained SDS-PAGE gel of human exocyst complex purified using eight different Twin-Strep tagged subunits (BV-SE1 to BV-SE8, see Table 1). (c) Coomassie-stained SDS-PAGE gel of human exocyst complex purified from insect cells co-infected with BV-2863 and BV-E1S547 (see Table 1). The exocyst complex was purified using Twin-Strep-tagged subunit EXOC5. (d) Electron micrograph of negative-stained recombinant human exocyst complex. The bar represents 100 nm. (e) Representative classes from 2D classification of recombinant human exocyst complex particles. (f) 3D reconstruction of recombinant human exocyst complex based nsEM data. (g) Coomassie-stained SDS-PAGE gel of the human dynactin complex purified by one-step strep-affinity purification. (h) Coomassie-stained 3–8% Native-PAGE gel of purified human dynactin complex after glycerol density gradient centrifugation purification. (i) Single-particle nsEM analysis of recombinant human dynactin complex with the representative raw micrograph (top) and 2D class averages (bottom). Scale bar, 50 nm. (j) Coomassie-stained SDS-PAGE gel of purified recombinant human COPI complex, human dynein complex, human CSN complex and human SCF complex.