A. Scheme depicting the PVD pathway involving biosynthesis, iron uptake and gene expression.

For details and explanations, refer to the “Introduction” section. The results obtained in this work on protein dynamics are indicated as follows: the stars in red, purple and blue indicate the proteins with rapid, moderate and slow dynamics, respectively. B. Fluorescence microscopy analysis of fluorescent fusions with, from left to right and up to down, OpmQ, FpvF, PvdQ, TonB, PvdT, PvdA and PvdS. Cells were grown twice in minimal medium, washed in minimal medium and spotted onto slides coated with agarose made up in minimal medium. Brightfield images, when available, are presented on the left. Due to low fluorescent signals, epifluorescence images of pvdS-yfp, mcherry-pvdT and mcherry-opmQ were recorded using a high sensitivity camera. Images of fpvF-mcherry in both epifluorescence (left panel) and TIRF (right panel) are shown. Epifluorescence images of pvdA-yfp, mcherry-pvdQ and pvdQ-mcherry are presented. For tonB-mcherry, from left to right, brightfield, epifluorescence and TIRF mode images are presented (scale bar 2 µm). For the fluorescence miscrocopy pictures of PAO1 strain harboring a plasmid encoding a cytoplasmic mCHERRY fluorescent protein expressed under the control of the pvdA promoter (PAO1(pMMB-mcherry)) see in Supplemental Materials (Figure 5-SM).如需了解详细信息与说明，请参阅「引言」部分。本研究中获得的蛋白质动力学相关结果标注如下：红色、紫色与蓝色星号分别代表具有快速、中等及慢速动力学特性的蛋白质。B. 荧光融合蛋白的荧光显微镜分析：按从左至右、从上至下的顺序，依次为OpmQ、FpvF、PvdQ、TonB、PvdT、PvdA与PvdS。菌体在基本培养基中传代两次，经基本培养基洗涤后，点样于以基本培养基配制的琼脂糖包被的载玻片上。若有明场图像，则置于左侧展示。由于荧光信号较弱，pvdS-yfp、mcherry-pvdT及mcherry-opmQ的落射荧光（epifluorescence）图像采用高灵敏度相机拍摄。本研究展示了fpvF-mcherry的落射荧光（左图）与全内反射荧光显微镜（Total Internal Reflection Fluorescence，TIRF，右图）成像结果。本次分析同时展示了pvdA-yfp、mcherry-pvdQ与pvdQ-mcherry的落射荧光图像。针对tonB-mcherry，按从左至右顺序依次展示明场、落射荧光及全内反射荧光显微镜成像（标尺为2 µm）。关于携带由pvdA启动子调控表达的胞质mCHERRY荧光蛋白质粒的PAO1菌株（PAO1(pMMB-mcherry)）的荧光显微镜图像，请参见补充材料（图5-SM）。