Progres image capture software

During the experiment, cyanobacterial samples (1 mL volume) were collected quantitively from cultures on day 6 of the experiment, and they were immediately preserved with Lugol’s iodine solution and used for microscopic analyses of trichome morphometry (thickness and length). The thickness of 10 and the length of 100 randomly selected trichomes were measured in each sample using an Axioskop 2 mot plus light microscope (Carl Zeiss AG, Oberkochen, Germany) equipped with a Jenoptik ProgRes Speed Xtcore3 digital camera and ProgRes image capture software (Jenoptik Optical Systems, Jena, Germany). Trichome thickness was measured in the middle part of the trichomes. Morphometric data were used to calculate the average trichome biovolume (mm³) following the formula for computing the volume of a cylinder, which corresponds to the geometric shape of A. gracile (Vadrucci et al., 2007 (link)). Trichome density in the samples was determined based on counting trichomes using a Beckman Coulter Cytomics FC 500 MPL flow cytometer (Beckman Coulter Life Sciences, Brea, CA, United States), and beads (C36950 Countbright Absolute Counting, Thermo Fisher Scientific, Waltham, MA, United States) were added as a counting reference (10 μL/1 mL of a sample). Trichome biovolume and density data were then used to calculate the biomass of A. gracile through the following formula:

LoVo, HT29 and RKO cells were cultured until 50-60% confluence before being fixed with 4% paraformaldehyde and being permeabilized with 0.3% Triton X-100. Cells were washed three times in phosphate-buffered saline (PBS) and incubated with PAK7, epithelial cadherin (E-cadherin) and neural cadherin (N-cadherin) (Abcam, UK) primary antibodies at 4°C overnight then, incubated with corresponding Alexa Fluor-conjugated secondary antibodies (Life Technologies, USA) at room temperature for 1 h, and mounted using ProLong® Gold Antifade Reagent with DAPI (Life Technologies, USA). Microscopic images of the cells were obtained using a Leica inverted fluorescence microscope (Leica Microsystems, Wetzlar, Germany) with ProgRes Image Capture Software (JENOPTIK Optical Systems, Jena, Germany) and a Leica Confocal LAS-AF SP5 System (Leica Microsystems, Germany).