Dataset Figure_4: Nuclear Whi5-GFP intensity versus time from repeated imaging of the same individual cells

Whi5-GFP intensity as a function of time for the same FOVs. Only small daughter cells in the asynchronous population at time point 0 were quantified as a function of time. The folder 'Image Files' contains raw image files for Whi5-GFP for FOV1,3,4,5 & 6 at each of the 6 time points (0, 20, 40, 60, 80, 100 min). The folder 'Excel_analysis_output' contains the output files for all five FOV (1,3,4,5,6) for 1. fovX_t0 or time 100_whi5_nadh_*.xlsx – at time 0 and time 100 minutes. These files correspond to the analysis of the background auto-fluorescence excited at 750 nm used to calculate size. The only relevant information from these analyses is the Cyto size (fL) column in tab 3 of each file 2. fovX_tX_whi5_*.xlsx Analysis of all time points and all FOV for Whi5-GFP intensities

Dataset Figure_5: Whi5-GFP intensity versus size and time in a synchronous G1 population

Whi5-GFP intensity as a function of time for the different FOV of elutriated cells. This dataset contains: 1. Raw TIF_images_NADH – These are autofluorescence images taken at each time point for the purposes of calculating cell size. Time points were every 10 minutes except at 30 minutes which had to be discarded due to poor focus. 2. Raw TIF_images_WHI5 – These raw image files correspond to images of Whi5-GFP excited at 1000 nm fpr all 11 time points of different FOV obtained at 3 different z positions (1-3) (0, -0.5 um, + 0.5 um). Whi5-GFP intensity values for each nucleus were taken for the z-position that gave the highest intensity for each nucleus. 3. NADHDATA_with_time plot.xlsx is the analysis of the raw images of auto-fluorescence exciting at 750 nm. The only relevant information for the Figure is in Colume C sheet 1. It is the cell area in total pixels. 4. Whi5Data_BestFocusPlanes_All_FOV.xlsx is the analysis of the Whi5-GFP images for Whi5-GFP intensity vs time and size for each time point (which corresponds to a different FOV)

Dataset Figure_6: Cln3 levels pulse prior to Start

Data and Matlab script 1. FLmean_Cln3_GLU: CSV file containing mean GFP concentration from Cln3-P2A-GFP for single daughter cells grown in glucose. Each cell is followed from birth until a few minutes after bud appearance. Data is organized by columns (one column corresponds to one cell). The first column is the vector of measurement time points (in minutes). 2. Vol_Cln3_GLU: CSV file containing cell volume for each of the daughter cells described above. Data is organized by columns (one column corresponds to one cell). The first column is the vector of measurement time points (in minutes). 3. bud_times: Excel file containing the time of bud appearance for each of the daughter cells described above 4. GP_FLtot_Cln3_GLU_example: Matlab script used to perform Gaussian process regression (see description in Methods) on the total GFP fluorescence for each of the daughter cells described above. The script produces single-cell Cln3 abundance data used to generate Fig. 5c of the main text and Extended Data Figure 4H. NOTE: in order to run, the Matlab script requires the installation of the GPML Matlab toolbox (freely available at http://www.gaussianprocess.org/gpml/code/matlab/doc/)