Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Proteomic analysis of FF samples revealed extracellular matrix (ECM) protein - vitronectin as the component responsible for FTE adhesion and spreading A. Experimental workflow of proteomics experiment. B. Venn diagram showing 14 common proteins identified between young and aged FF samples from proteomics analysis. Common proteins are listed with vitronectin (highlighted in red). C. Representative immunoblot for vitronectin expression in 3 young (Y1–Y3) and 3 aged (A1-A3) FF samples. FF samples (5 μg) and recombinant vitronectin protein (0.01 μg, 0.1 μg, 1 μg) diluted with lysis buffer were used for immunoblotting. Arrow represents the band analyzed for vitronectin. Ponceau staining was used for loading control.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques: Western Blot, Expressing, Recombinant, Lysis, Staining, Control

Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Vitronectin in FF aids in FTE adhesion and spreading A. Representative brightfield images of FT190 cells seeded on ULA plates coated with FF sample (400 μl) and recombinant vitronectin protein (1 μg/well) for 4 h. Wells were washed with 1X PBS and FT190 cells were seeded on the coated plates. Images were acquired after 24 h. Scale bar = 200 μm. B. Cell proliferation was measured using an SRB assay to measure cell viability of FT190 cells on FF and vitronectin coated ULA plates. C. Representative Z stack images acquired by confocal microscopy of FT190 spheroids (labelled with Cell tracker 594) on NOF151 cells (labelled with Cell tracker 488) with FF samples and recombinant vitronectin protein (1 μg) (24 h). Scale bar = 200 μm. D. A maximum intensity projection re-construction of a three-dimensional FTE spheroid (labelled with Cell tracker 594) optical data stack over the surface of NOF151 cells (labelled with Cell tracker 488) treated with FF samples (Y1, A1) and vitronectin (1 μg). Spheroids were imaged with a 10× objective and re-constructed using the Imaris software. E. Representative images acquired by confocal microscopy showing a side and top projection of the FT190 spheroids on NOF151 cells with FF samples and recombinant vitronectin protein (1 μg) (24 h). Scale bar = 100 μm. F. Spheroid area (with and without FF/vitronectin) was determined by quantification of red fluorescent intensity using a polygon area measurement tool of the Imaris software.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques: Recombinant, Sulforhodamine B Assay, Confocal Microscopy, Construct, Software

Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Primary antibodies.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques:

Journal: NPJ Biofilms and Microbiomes

Article Title: Clostridium difficile -derived membrane vesicles promote fetal growth restriction via inhibiting trophoblast motility through PPARγ/RXRα/ANGPTL4 axis

doi: 10.1038/s41522-024-00630-5

Figure Lengend Snippet: A The protein expression levels of β-actin, FN1 and VTN were detected by western blotting. B The mRNA expression levels of FN1 and VTN was detected by qRT-PCR. C Adhesion ability of the HTR-8/SVneo cells treated with C. difficile MVs 5 μg/ml was detected by adhesion experiment. D Migration ability of the HTR-8/SVneo cells treated with C. difficile MVs 5 μg/ml for 24 h was detected by wound-healing assay. E Transwell assay was used to analyze the migration and invasion capabilities of HTR-8/SVneo cells treated with C. difficile MVs 5 μg/ml for 24 h. Data are represented as mean ± SD including at least three replicates. Data was analyzed using t -test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Membranes were incubated with the following antibodies: FN1 (1:2000, 15613-1-AP, Proteintech), VTN (1:10000, 66398-1-Ig, Proteintech), PPARγ (1:1000, 60127-1-Ig, Proteintech), and β-actin (1:10000, 66009-1-Ig, Proteintech).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Migration, Wound Healing Assay, Transwell Assay

Journal: NPJ Biofilms and Microbiomes

Article Title: Clostridium difficile -derived membrane vesicles promote fetal growth restriction via inhibiting trophoblast motility through PPARγ/RXRα/ANGPTL4 axis

doi: 10.1038/s41522-024-00630-5

Figure Lengend Snippet: A The inhibitory effect of C. difficile MVs on HTR-8/SVneo cell adhesion was mitigated by T0070907 and PPARγ shRNA. B Down-regulation of FN and VTN by C. difficile MVs was eliminated by T0070907 and PPARγ shRNA in HTR-8/SVneo cells. C , D The inhibition of C. difficile MVs on the migration and invasion of HTR-8/SVneo cells was alleviated by T0070907 and PPARγ shRNA. Data are represented as mean ± SD including at least three replicates. Data was analyzed using ANOVA with Tukey’s post hoc test. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Membranes were incubated with the following antibodies: FN1 (1:2000, 15613-1-AP, Proteintech), VTN (1:10000, 66398-1-Ig, Proteintech), PPARγ (1:1000, 60127-1-Ig, Proteintech), and β-actin (1:10000, 66009-1-Ig, Proteintech).

Techniques: shRNA, Inhibition, Migration

Journal: NPJ Biofilms and Microbiomes

Article Title: Clostridium difficile -derived membrane vesicles promote fetal growth restriction via inhibiting trophoblast motility through PPARγ/RXRα/ANGPTL4 axis

doi: 10.1038/s41522-024-00630-5

Figure Lengend Snippet: A Protein expression levels of PPARγ, ANGPTL4 and RXRα were elevated in the FGR placenta, and the levels of VTN and FN1 were decreased. B The mRNA expression level of PPARγ was significantly increased in the placenta of FGR. Data are represented as mean ± SD including at least three replicates. Data was analyzed using t -test. * P < 0.05; ** P < 0.01; *** P < 0.001. C – G Correlations between the relative mRNA expression level of PPARγ and patient clinical parameters. Spearman’s correlation test was used for statistical analysis. H The schematic diagram of C. difficile and its MVs involvement in FGR.

Article Snippet: Membranes were incubated with the following antibodies: FN1 (1:2000, 15613-1-AP, Proteintech), VTN (1:10000, 66398-1-Ig, Proteintech), PPARγ (1:1000, 60127-1-Ig, Proteintech), and β-actin (1:10000, 66009-1-Ig, Proteintech).

Techniques: Expressing

Journal: NPJ Biofilms and Microbiomes

Article Title: Clostridium difficile -derived membrane vesicles promote fetal growth restriction via inhibiting trophoblast motility through PPARγ/RXRα/ANGPTL4 axis

doi: 10.1038/s41522-024-00630-5

Figure Lengend Snippet: Primers used in qRT-PCR

Article Snippet: Membranes were incubated with the following antibodies: FN1 (1:2000, 15613-1-AP, Proteintech), VTN (1:10000, 66398-1-Ig, Proteintech), PPARγ (1:1000, 60127-1-Ig, Proteintech), and β-actin (1:10000, 66009-1-Ig, Proteintech).

Techniques: Sequencing