penta mutant sortase a esrta  (Addgene inc)

Journal: Scientific Reports

Article Title: inhibitory effect of hyperin on Staphylococcus aureus pathogenicity though interactions with sortase A and sortase B

doi: 10.1038/s41598-025-23458-1

Figure Lengend Snippet: Hyperin reduces biofilm formation, adhesion and cytotoxicity. ( a ) The absorbance at 600 nm of S. aureus USA300 when it was cocultured with various concentrations of hyperin. ( b ) Biofilm formation of S. aureus USA300 after treatment with different concentrations of hyperin. S. aureus USA300 was cultured with different concentrations of hyperin for 24 h, the samples were stained with 0.2% crystal violet, and the absorbance at 570 nm was measured after treatment with 35% glacial acetic acid. Data are shown as the mean with the SD, ** indicates p ≤ 0.01. ( c ) Adhesion of S. aureus USA300 to lung epithelial cells when different concentrations of hyperin were added. A549 cells were treated with S. aureus USA300 and different concentrations of hyperin for 1.5 h, after which the cells were washed, harvested, plated onto LB agar medium and cultured. Data are shown as the mean with the SD, ** indicates p ≤ 0.01. ( d ) Survival rate of A549 cells treated with hyperin. The data are shown as the mean and SD. ( e ) LDH levels in A549 cells and the survival of cells determined by live/dead staining ( f ). A549 cells were treated with S. aureus USA300 and different concentrations of hyperin for 6 h. The LDH levels in the supernatant were measured with an LDH kit, and the cells were stained with live/dead reagents. Data are shown as the mean with the SD, ** indicates p ≤ 0.01. The bar represents 50 μm. ( g ) The expression levels of SrtA and ClfA in S. aureus USA300 treated with different concentrations of hyperin. S. aureus USA300 was cocultured with different concentrations of hyperin, after which the bacteria were harvested by centrifugation and treated with SDS-PAGE loading buffer. The proteins were separated on a 10% gel and identified by specific antibodies. ( h ) Quantitative analysis of the expression levels of SrtA and ClfA ( i ). The analysis was performed with Image J 1.54 g. The data are shown as the mean and SD.** indicates p ≤ 0.01.

Article Snippet: ClfA antibody (Cusabio, CSB-PA692021ZA01FLB, 1:1000), and SrtA antibody (cusabio, CSB-EP3093FLF, 1:3000).

Techniques: Cell Culture, Staining, Expressing, Bacteria, Centrifugation, SDS Page

Journal: Scientific Reports

Article Title: inhibitory effect of hyperin on Staphylococcus aureus pathogenicity though interactions with sortase A and sortase B

doi: 10.1038/s41598-025-23458-1

Figure Lengend Snippet: Hyperin reduces biofilm formation, adhesion and cytotoxicity. ( a ) The absorbance at 600 nm of S. aureus USA300 when it was cocultured with various concentrations of hyperin. ( b ) Biofilm formation of S. aureus USA300 after treatment with different concentrations of hyperin. S. aureus USA300 was cultured with different concentrations of hyperin for 24 h, the samples were stained with 0.2% crystal violet, and the absorbance at 570 nm was measured after treatment with 35% glacial acetic acid. Data are shown as the mean with the SD, ** indicates p ≤ 0.01. ( c ) Adhesion of S. aureus USA300 to lung epithelial cells when different concentrations of hyperin were added. A549 cells were treated with S. aureus USA300 and different concentrations of hyperin for 1.5 h, after which the cells were washed, harvested, plated onto LB agar medium and cultured. Data are shown as the mean with the SD, ** indicates p ≤ 0.01. ( d ) Survival rate of A549 cells treated with hyperin. The data are shown as the mean and SD. ( e ) LDH levels in A549 cells and the survival of cells determined by live/dead staining ( f ). A549 cells were treated with S. aureus USA300 and different concentrations of hyperin for 6 h. The LDH levels in the supernatant were measured with an LDH kit, and the cells were stained with live/dead reagents. Data are shown as the mean with the SD, ** indicates p ≤ 0.01. The bar represents 50 μm. ( g ) The expression levels of SrtA and ClfA in S. aureus USA300 treated with different concentrations of hyperin. S. aureus USA300 was cocultured with different concentrations of hyperin, after which the bacteria were harvested by centrifugation and treated with SDS-PAGE loading buffer. The proteins were separated on a 10% gel and identified by specific antibodies. ( h ) Quantitative analysis of the expression levels of SrtA and ClfA ( i ). The analysis was performed with Image J 1.54 g. The data are shown as the mean and SD.** indicates p ≤ 0.01.

Article Snippet: ClfA antibody (Cusabio, CSB-PA692021ZA01FLB, 1:1000), and SrtA antibody (cusabio, CSB-EP3093FLF, 1:3000).

Techniques: Cell Culture, Staining, Expressing, Bacteria, Centrifugation, SDS Page