In all growth assays, bacteria were subcultured in 50 ml of growth media for a starting OD600 of 0.05.
After 1 h incubation for cell adhesion, 2 mL of growth medium was added into each well and plates were incubated at 37 °C and 5% CO2.
Viable cells were plated at 500,000 cells/cm2 in 40 mL of growth media ±10 ng/mL bFGF (Austral Biologicals, San Ramon, CA, cat# GF-030-5 GF-030-5 GF-030-5e flask (Nunclon®, Naperville, IL, cat# 178883).
The cells were washed twice with phosphate-buffered saline (PBS) and resuspended in 10 mL of growth medium a-MEM (10% fetal bovine serum (FBS), 100 mg/mL streptomycin, 100 U/mL penicillin, 2 mmol/L l-glutamine and 25 ng/mL amphotericin B).
After cell seeding (control, with 50 µM blebbistatin and with siRNA FAK sc-29310 Santa Cruz Biotechnology transfected with Neon Transfection System, ThermoFisher) onto double rigidity hydrogels, samples were transferred to microscope culture chamber (37 °C, 5% CO2) and gently submerged in 5 ml of growth media.
The hydrogen autotrophic fermentation experiment was carried out in a 1 L serum bottle containing 400 ml of growth medium.
Each sterilized chamber containing arsenopyrite was filled to half capacity with a total of 210 ml of growth medium.
The vials contained 50 ml of growth medium and the initial glucose concentration was in the range of 9 50 g L−1.
Next, 5 mL of growth medium (1500 cfu containing Luria Bertani medium) for each bacterial species was taken in sample-contained tube.
The growth experiment was conducted in dark (12 h) and light (12 h) cycle conditions using 300 mL of growth media in 500 mL conical flasks.
Cells were seeded in 75 cm2 flasks with 15 ml of growth medium, unless otherwise mentioned.
