Fig. 3: Reconstitution of FtsZ-FtsA mesh and Min wave regulated FtsZ-ring structures via cell-free expression.

a Schematic illustration of PURE cell-free co-expression of FtsA, MinD, and MinE with purified FtsZ-Alexa488 and mCherry-MinC. b 3D max projection of FtsZ-Alexa488 mesh structures inside vesicles via FtsA expression (after 1 h of expression with 5 nM of ftsA template). Scale bar: 10 μm. c Regulation of FtsZ localization by Min waves. Left: 3D max projection of the transition of FtsZ structures and Min wave patterns by co-expression of FtsA, MinD, and MinE within vesicles (2 nM ftsAopt and 2 nM minDEoperon templates). Starting from no patterns, an FtsZ mesh develops on the membrane, which is then instantly condensed into an FtsZ-ring structure by the later onset of pole-to-pole oscillations of Min waves, through antagonistic localization of Min and FtsZ proteins. Right: Kymographs of the pattern transition of Min waves and FtsZ structures. Recording started after 5 min of cell-free expression. mCherry-MinC and FtsZ-Alexa488 are indicated in magenta and green, respectively. d Time-lapse images of FtsZ-ring like structures induced by pole-to-pole oscillations of Min waves. Experimental condition is the same as Fig. 3c and recording started after 30 min of cell-free expression. Scale bar: 5 μm.