The biofilm formation of wild type strain PAO1 with or without the extracts of A. pilosa or wogonin was tested according to the previous method [48 (link)]. The strain with different concentrations of the extracts of A. pilosa or wogonin was added in sterile test tubes and incubated at 37 °C until the biofilm was formed. Followed by removal of non-adherent cells from the tubes and gentle rinsing three times with sterile PBS, biofilm biomass was evaluated by crystal violet staining. The attached crystal violet was washed with anhydrous ethanol and its absorbance was measured at OD570 and analyzed.
In order to determine the effects of wogonin on the formed biofilm, after PAO1 was grown in a 96-well polyvinyl chloride plate (Corning/Costar, Corning, NY, USA) at 37 °C for 10 h, the culture was removed and new LB broth with 15 or 30 μg/mL wogonin was added and incubated at 37 °C overnight. The biofilm biomass was evaluated by crystal violet staining as the above-mentioned. Each samples had 12 replicates.
In order to determine the effects of wogonin on the formed biofilm, after PAO1 was grown in a 96-well polyvinyl chloride plate (Corning/Costar, Corning, NY, USA) at 37 °C for 10 h, the culture was removed and new LB broth with 15 or 30 μg/mL wogonin was added and incubated at 37 °C overnight. The biofilm biomass was evaluated by crystal violet staining as the above-mentioned. Each samples had 12 replicates.