Skip to main content
. 2019 Jan 23;4(1):e00689-18. doi: 10.1128/mSphere.00689-18

FIG 1.

FIG 1

The general muscarinic receptor agonist pilocarpine hydrochloride inhibits Candida albicans biofilm formation in vitro. (A and B) Biofilm biomass was assessed using the crystal violet assay after C. albicans was cultured for 24 h in RPMI 1640 containing different concentrations of SIBm (0 to 10 mM) (A) and PHCl (0 to 50 mM) (B). Data are expressed as raw OD595 values, and the bars represent the mean values (plus standard deviations [SD] [error bars]) from triplicate wells of six independent experiments (n = 6). (C and D) Biofilm metabolic activity was assessed using the XTT assay after C. albicans was cultured for 24 h in RPMI 1640 containing different concentrations of SIBm (0 to 10 mM) (C) and PHCl (0 to 50 mM) (D). Data are expressed as percent metabolic activity compared to untreated controls (0 mM PHCl), and the bars represent the mean values (+SD) from triplicate wells of six independent experiments (n = 6). For panels A to D, values that are significantly different compared to the control values (0 mM SIBm or PHCl) are indicated by asterisks as follows: *, P < 0.05; **, P < 0.01; ***, P < 0.001. (E and F) The effect of PHCl on the permeability of the C. albicans cell wall was investigated using a propidium iodide (PI) uptake (E) and ATP release assay (F). For the PI uptake assay, data are shown as fluorescence intensity units, and the bars represent the mean values (+SD) from triplicate wells of three independent experiments (n = 3). Heat-killed (HK) and chlorhexidine (CHX) (0.2%)-treated C. albicans were included as positive controls, and cells in RPMI 1640 alone were included as a negative control. For the ATP release assays, data are shown as nanomolar concentrations of ATP release, and the bars represent the mean values (+SD) from triplicate wells of three independent experiments (n = 3). Chlorhexidine (0.2%)- and fulvic acid (2.0%)-treated C. albicans cells were included as positive controls, and cells in RPMI 1640 alone were included as a negative control. *, P < 0.05; **, P < 0.01 compared to the control (cells in RPMI 1640 alone).