Shikonin Inhibits Candida albicans Biofilms via the Ras1-cAMP-Efg1 Signalling Pathway

Figures & data

Table 1 Primers Used in This Study

Number	Primer	Sequence
1	ECE1-F	GCTGGTATCATTGCTGATAT
	ECE1-R	TTCGATGGATTGTTGAACAC
2	HWP1-F	TGGTGCTATTACTATTCCGG
	HWP1-R	CAATAATAGCAGCACCGAAG
3	ALS3-F	CTAATGCTGCTACGTATAATT
	ALS3-R	CCTGAAATTGACATGTAGCA
4	RAS1-F	GGCCATGAGAGAACAATATA
	RAS1-R	GTCTTTCCATTTCTAAATCAC
5	CYR1-F	TTCATCAGGGGTTATTTCAC
	CYR1-R	CTCTATCAACCCGCCATTTC
6	EFG1-F	TATGCCCCAGCAAACAACTG
	EFG1-R	TTGTTGTCCTGCTGTCTGTC
7	TEC1-F	AGGTTCCCTGGTTTAAGTG
	TEC1-R	ACTGGTATGTGTGGGTGAT
8	18S rRNA-F	AATTACCCAATCCCGACAC
	18S rRNA-R	TGCAACAACTTTAATATACGC

Abbreviations: F, forward primer; R, reverse primer.

Figure 1 Morphology alteration of C. albicans SC5314 biofilms after treated by 8 μg/mL SK.

Notes: The inset in the 3500×, 2000×, 1000× panels show the area that was magnificated.

Figure 2 Inhibitory effect of different concentrations of SK on cell adhesion of C. albicans SC5314.

Notes: (A) Control; (B) 4 μg/mL SK; (C) 8 μg/mL SK; (D) 16 μg/mL SK compared with control, **P < 0.01.

Figure 3 Effects of different concentrations of SK on cell surface hydrophobicity (CSH) of C. albicans SC5314 biofilms.

Notes: Compared with control, *P < 0.05, **P < 0.01.

Figure 4 Changes in biofilms related genes of RAS1-cAMP-EFG1 pathway expression after treated by SK (8 μg/mL) for C. albicans SC5314.

Notes: The mRNA levels were normalized on the basis of their 18S rRNA levels, gene expression was calculated as the fold increase relative to the control group.

Figure 5 Effects of SK on the production of cAMP protein of C. albicans SC5314 biofilms.

Notes: Compared with control, **P < 0.01.

Figure 6 Exogenous cAMP reverts the inhibitory effect of SK on C. albicans SC5314 biofilms. Biofilm formation was evaluated by XTT reduction assay, and the results were presented as the percentage compared to the control biofilms formed without drug treatment.

Notes: Compared with different concentrations of SK single use groups, *P < 0.05, **P < 0.01.

Data Sharing Statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.