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: bioRxiv

Article Title: A circular engineered sortase for interrogating histone H3 in chromatin

doi: 10.1101/2024.09.10.612318

Figure Lengend Snippet: ( A ) Workflow for isolating tandem mass tagged histone H3 tails with cW11 sortase. ( B ) SDS-PAGE of 14-hour sortase reaction in a nuclear acid extract. ( C ) Tris-Tricine PAGE of histone H3(1-34) synthetic standards (1: 140 ng; 2: 420 ng) and TCA precipitated H3(1-32)-TMT from the acid extract reaction; bands between 25 and 37 kDa are histone H1. ( D ) Structure of the TMT-labelled oligoglycine peptide illustrating sortase reactive GGG, charge carrying K and carboxamide, and TMT labelled aminoalanine. ( E ) Unique proteoforms detected in each 6plex sample and shared between the two samples. ( F ) Quantifiable proteoforms detected in each 6plex sample and shared between the two samples. Volcano plot of significant (p<0.05) log2-fold change and overall abundance of H3 proteoforms in HEK293T cells following treatment with ( G ) LSD1/HDAC1/CoREST complex-specific inhibitor Corin, or ( H ) pan class one HDAC inhibitor MS275.

Article Snippet: The development of cW11 builds on the earlier work of Schwarzer and colleagues who reported F40 sortase as a tool for histone H3 semisynthesis.

Techniques: SDS Page

Journal: bioRxiv

Article Title: A circular engineered sortase for interrogating histone H3 in chromatin

doi: 10.1101/2024.09.10.612318

Figure Lengend Snippet: Single, combinatorial and asymmetric modifications prepared by sortase nucleosome ligation (ubiquitin structure PDBID: 1UBQ). Isolated yields (bold) and yields estimated by area under the curve (parenthetical) are reported as a value (n = 1), range (n = 2) or mean with standard deviation (n ≥ 3).

Article Snippet: The development of cW11 builds on the earlier work of Schwarzer and colleagues who reported F40 sortase as a tool for histone H3 semisynthesis.

Techniques: Ligation, Ubiquitin Proteomics, Isolation, Standard Deviation

Journal: bioRxiv

Article Title: A circular engineered sortase for interrogating histone H3 in chromatin

doi: 10.1101/2024.09.10.612318

Figure Lengend Snippet: (Top) Asymmetric modifications prepared by sortase nucleosome ligation. Isolated yields (bold) and yields estimated by area under the curve (parenthetical) are reported as a value (n = 1), range (n = 2) or mean with standard deviation (n ≥ 3). (Bottom left) Western blot characterization of 147 bp nucleosomes: ( 1 ) H3 (aa33-135) starting material; ( 2 ) asymmetric H3K27ac & H3 (aa33-135) intermediate; ( 3 ) asymmetric H3K27ac & H3K9ac/K14ac/K18ac/K23ac product. (Bottom right) Deconvoluted mass spectrum of 147 bp asymmetric H3K27ac & H3K9ac/K14ac/K18ac/K23ac nucleosome.

Article Snippet: The development of cW11 builds on the earlier work of Schwarzer and colleagues who reported F40 sortase as a tool for histone H3 semisynthesis.

Techniques: Ligation, Isolation, Standard Deviation, Western Blot