Fig. 2: Identification of a cytosolic PodJ fragment as the opposite-pole oligomer.
From: Construction and phenotypic classification of synthetic dual-pole Escherichia coli cells
a Domain organization of the PodJ variants tested in this study (from top to bottom: full-length PodJ, PodJCC1−3, IDR, TM, and PodJCC1−3, IDR). “CC”, “IDR,” and “TM” denote the coiled-coil region, intrinsically disordered region, and transmembrane domain, respectively. b Circuit diagrams of the sfGFP-fused PodJ variants expressed from the IPTG-inducible Ptac promoter. c Images showing the bipolar localization pattern of sfGFP-fused PodJ variants, induced by 250 μM IPTG at 37 °C for 1 h. Scale bars: 1 μm. d Circuit diagrams illustrating the co-expression of sfGFP-PodJ variants with mRFP-PopZ. The sfGFP-PodJ variants were expressed under the Ptac promoter, while mRFP-PopZ was expressed from the Anderson promoter J23116. e Images showing mRFP-PopZ expression patterns with or without co-expressed sfGFP-PodJ variants. sfGFP-PodJ variants were induced by 500 μM IPTG at 28 °C for 2 h. Scale bars: 1 µm. f Quantification of PodJ variant co-localization with PopZ in (e). n = 60, 79, and 74 for cells expressing PodJ, PodJCC1−3, IDR, TM, and PodJCC1−3, IDR, respectively. The center line represents the median, box limits indicate the upper and lower quartiles, and whiskers extend to 1.5× the interquartile range. Statistical differences were determined using a two-tailed Student’s t test; **** indicates p < 0.0001.
