Fig. 4
From: Protein polarization driven by nucleoid exclusion of DnaK(HSP70)–substrate complexes
LDCs appear and remain at the bacterial pole. a, b Distribution of LDCs along the cell major axis. Green: Civ; cyan: DAPI. a (left) Representative micrographs of bacteria with DAPI and Ci fluorescence, shown are cell outline, mid-line and foci detected using MicrobeJ software. b Mean ± SEM of the fluorescence intensity relative to the cell major axis (106 bacteria, N = 3). c–i Live cell fluorescence microscopy analysis of assembly and disassembly of LDCs. c, f Stills from time-lapse series, with the elapsed time indicated in min. d, g Tracks of LDCs’ peak intensity of foci along the bacterial major axis (shown are representative tracks). Dashed line indicates limit of detection. e Traces: fluorescence intensity of single LDCs corrected for initial intensity plotted over time (Shown are traces from six individual bacteria). h Mean ± SEM of LDC fluorescence intensity normalized to initial fluorescence (% Intensity; 1 field of view (six bacteria), N = 3). Holm-Sidak test, statistical significance relative to untreated bacteria, asterisk: p < 0.05. i Mean ± SEM of the number of foci in the presence or absence (CTRL) of DNP at the onset of analysis (T0) or following 60 min incubation (TF) (foci were counted in 278, 352, 1096 and 486 bacteria from CTRL and DNP (T0), CTRL and DNP (TF), respectively; N = 3). Statistical significance determined with Kruskal–Wallis test followed by Dunn’s post hoc analysis, four asterisks: p < 10−4. d, e MC4100/pCiv. f–i MC4100/pSUCi. Arrows: LDCs. Scale bar = 2 µm
