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. 2017 Feb 24;7:43306. doi: 10.1038/srep43306

Figure 4. Effect of FtsW, PBP3 and the FtsW-PBP3 complex on the polymerization of lipid II by PBP1b and the mobility of lipid II (LII) analyzed by (SDS- or native) PAGE.

Figure 4

(A and B) radioactive lipid II revealed by storage phosphor imaging using Typhoon trio+ (GE Health Care). (A) SDS-PAGE analysis of PBP1b (1b) or PBP1bTP* mutant (1b*) (both at 100 nM) reaction products in the presence of PBP3 (3) (3 μM), FtsW (W) (3 μM), and the FtsW-PBP3 (W3) complex (3 μM). (B) effect of FtsW (3 μM) and the FtsW-PBP3 complex (3 μM) in the absence of PBP1b on the mobility of lipid II; buffer with excess detergent used as control. (C) Co-migration of FtsW and NBD-lipid II in SDS-PAGE. (D) analysis of NBD-lipid II (10 μM) migration by native-PAGE in the absence and the presence of FtsW, the FtsW-PBP3 complex or MurJ (all used at 5 μM). (C and D) fluorescent lipid II was first revealed by fluorescence imaging using Typhoon trio+, then the gels were stained with Coomassie blue.