Additional file 1 of Mechanical force-induced morphology changes in a human fungal pathogen
Creators: name: Puerner, Charles
name: Kukhaleishvili, Nino
name: Thomson, Darren
name: Schaub, Sebastien
name: Noblin, Xavier
name: Seminara, Agnese
name: Bassilana, Martine
name: Arkowitz, Robert A.
Publisher: name: figshare
Year: 2020
Subjects: subject: Biophysics
subject: 29999 Physical Sciences not elsewhere classified
subject: FOS: Physical sciences
subject: FOS: Physical sciences
subject: Microbiology
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Cell Biology
subject: Biotechnology
subject: 59999 Environmental Sciences not elsewhere classified
subject: FOS: Earth and related environmental sciences
subject: FOS: Earth and related environmental sciences
subject: Ecology
subject: Science Policy
subject: Cancer

description: Additional file 1: Figure S1. Strain versus stress dependence of PDMS. Analyses carried out using a Viscoanalyzer, with oscillation at 10 Hz of PDMS at cross-linker ratio of 40:1. Figure S2. Invasive growth and penetration into adjacent chamber in PDMS of different stiffness. DIC time-lapse experiments at indicated PDMS:cross-linker ratio and measured stiffness (Young’s modulus). The adjacent chamber is highlighted with a dotted yellow line and deformation of this chamber lasted ~40 min with 40:1 PDMS ratio and 80-90 min for the two stiffer PDMS substrates. Figure S3. The shape of filament tip is not substantially altered during invasive growth in PDMS. A) Radius of curvature over time is constant in surface and invasively growing cells. Radius of curvature with an arc of ± 90° or ± 45° at the filament tip. B) Shape of filament tip of surface growing cells over time. Cells were grown on PDMS (30:1; 250 kPa) and 31 × 5 min GFP sum projections were analyzed. Radius of curvature with ± 45° by indicated open lines and ± 90° indicated by solid lines. Figure S4. Cells confined within a stiff PDMS chamber have reduced filament extension rates. A) Constricted growth within a PDMS chamber. Typical time-lapse experiment using 160 kPa PDMS, with DIC images every 5 min shown. B) Filament extension rate within a stiff chamber is not linear. Filament length was determined from images every 5 min for ~ 2 h and GFP sum projections (n = 9 cells). C) Filament extension rate is substantially reduced as chamber fills up. Initial (filament length 10-20 μm) and final (filament length &gt; 20 μm) extension rates were determined from fits to 6 × 5 min GFP sum projections. (colors represent individual cells). Bars indicate SD and **** p &lt; 0.0001. Figure S5. Distribution of active Cdc42 is not altered during invasive growth. A) Schematic indicating fluorescence signal over the filament long axis. Quantitation of slope of Gaussian farthest from tip in red (Max Slope, in relative units), distance maximum signal to tip (xmax in μm), and half width half max of the Gaussian farthest from tip in red (xSpread-xmax), i.e. the signal spread (Spread in μm). Signal is denoted by I and distance from tip by x. B) Distribution of active Cdc42 during surface and invasive filamentous growth. Experiment described in Figure 11a and 11b with the mean signal for each cell (colors represents individual cells), normalized to the mean signal for tip Cdc42•GTP in surface growing cells. Bars indicate SD. C) Distribution of active Cdc42 is not altered upon invasive growth. Relative maximum slope (left), distance from maximum signal to the tip (middle) and spread of signal (right) determined from 6-8 cells, using tailor-made Matlab program. Bars indicate SD; surface and invasive cells were not significantly different. D) Apical and subapical active Cdc42 signals are stable over time. Relative signals from apical and subapical region of sum projections, normalized to maximum invasive subapical signal.