Fig. 1: Identification of Glp as a member of the corynebacterial divisome.
a, The core interactome of SepF, including proteins recovered from three independent co-IP experiments using different strains/antibodies: Cglu/α-SepF, Cglu_SepF-Scarlet/α-SepF and Cglu_SepF-Scarlet/α-Scarlet. The square size for each interactor is proportional to its enrichment in the interactome compared to the proteome (Supplementary Table 1), and the lines indicate either direct or indirect interactors. b, Glp depletion and complementation. Representative images in phase contrast (PC) and membrane staining for indicated strains. c, Left: frequency histogram showing the number of septa per cell for the different strains, calculated from n cells imaged (indicated in the figure) from three independent experiments for each strain (Cglu_Δglp, n = 873, 1,538 and 840; Cglu, n = 718, 1,468 and 1,223; Cglu_Δglp + Glp, n = 2,465, 1,169 and 1,297; Cglu_Δglp + mNeon-Glp, n = 1,641, 1,311 and 1,801); open circles represent the corresponding data points; mean ± s.d.; Cohen’s d (see Methods for interpretation of values), from top to bottom: (***d = 1.57, P = 0), (***d = 1.84, P = 0), (***d = 1.6, P = 0). Middle and right: violin plots showing the distribution of cell length (Cohen’s d, from top to bottom: (***d = 1.76, P ~ 0), (nsd = 0, P = 0.95), (nsd = 0.29, P = 3.78 × 10−37), (nsd = 0.27, P = 2.59 × 10−39)) and cell width (Cohen’s d: (****d = 2.21, P ~ 0), (nsd = 0.25, P = 4.13 × 10−25), (nsd = 0.38, P = 9.65 × 10−74), (nsd = 0.08, P = 1.74 × 10−12)); the box indicates the 25th to the 75th percentile, the mean and the median are indicated with a dot and a line in the box, respectively. d, Ethambutol sensitivity assay. BHI overnight cultures of Cglu and Cglu_Δglp complemented with the empty plasmid or mNeon-Glp were normalized to an OD600 of 0.5, serially diluted 10-fold and spotted onto BHI agar medium with or without 1 μg ml−1 ethambutol. e, Left: localization of mNeon-Glp in Cglu. Representative images in PC, membrane staining and mNeon-Glp fluorescent signals. The arrow indicates the Glp localization before septum formation. Right: heat map representing the localization pattern of mNeon-Glp; 3,879 cells were analysed, from triplicate experiments. Scale bars, 5 μm.
