Extended Data Fig. 1: Processing raw single-cell data: smoothing single-cell traces of volume and sfGFP fluorescence, obtaining the production rate of sfGFP expressed from the heterologous promoter tetO7, with and without accounting for sfGFP maturation.
(a,b) Single-cell volume (a) and sfGFP fluorescence (b) during an individual cell cycle. We show raw data with asterisks when corresponding to the daughter cell, triangles when corresponding to the mother cell and circles when corresponding to the whole cell. The vertical lines denote major cell cycle events: mitotic exit (ME), START and budding (BUD). The curves show the LOWESS smoothing with adjacent smoothed values (at the same time points as the raw data) connected with a line. a: the discontinuity in the values that should immediately follow mitotic exits due to cytokinesis is tackled by vertically moving down the data before the first mitotic exit and up the data after the second mitotic exit. b: we measured sfGFP fluorescence only in the mother cell and assumed that it is the same in the whole cell. (c–e) Single-cell sfGFP abundance (c), sfGFP production rate calculated with (d) and without (e) accounting for the maturation of the fluorescent protein (assumed maturation half-time 6 minutes, first-order maturation kinetics). The curves show linearly connected values corresponding to the same time points as the raw data. c: values are the product of the smoothed cell volume (a) and smoothed sfGFP fluorescence (b). d: values are a linear combination of the first and second derivatives of sfGFP abundance (c). e: values are the first derivative of sfGFP abundance (c). (f, g) The production rate of sfGFP presented as a summary of 38 individual cell-cycle traces. The sfGFP production rate was computed without accounting for the maturation of this fluorescent protein. Values from (e) and other 37 cell cycles correspond to the markers. The plots were built analogously to Fig. 1a, b.
