Quantification of sfGFP-MinD, MinD, and MinE. (A) Validation of the ability of the anti-MinD antiserum to detect sfGFP-MinD and MinD in cell lysates alongside purified His6x-sfGFP-MinD and Trx-His6x-MinD. The blot-purified antiserum was used at a 1:100 dilution. (B) Validation of the anti-MinE antiserum ability to detect MinE in cell lysates alongside MinE-His6x. The blot-purified antiserum was used at a 1:100 dilution. (C and D) An example of the western blots used to determine the concentrations of MinD in the cell lysates of the W3110 and FW1541 strains, respectively. Three independent cultures (sample repeats n = 3) were collected for each western blot, and the experiment was repeated at least twice for each strain. (E and F) An example of the western blots used to determine the concentrations of MinE in the cell lysates of the W3110 and FW1541 strains, respectively. Three independent cultures (sample repeats n = 3) were collected for each western blot, and the experiment was repeated five times for each strain. In C–F, serial dilutions of purified Trx-His6x-MinD, His6x-sfGFP-MinD, or MinE-His6x were applied to generate a calibration curve for interpolating the amount of MinD, sfGFP-MinD, or MinE in the sample by linear regression. Source data are available for this figure: SourceData FS1.
