fluorescein labeled lectin  (Vector Laboratories)

Journal: PLOS One

Article Title: DJ-1 alleviates high glucose-induced podocyte injury via activating ERK1/2 signaling

doi: 10.1371/journal.pone.0346714

Figure Lengend Snippet: (A) Representative Western blot images and quantitative statistical bar graphs of protein expression levels of DJ-1 in HPCs treated with mannitol (MG) or D-glucose were analyzed by Western blotting analysis and Image J software, respectively. (B) The mRNA levels of DJ-1 in HPCs treated with MG or D-glucose were analyzed by qRT-PCR analysis. (C) The cell viability was measured by CCK-8 assays in HPCs treated with MG, LG, HG and HG + DJ-1, respectively. (D) The cell viability was analyzed by EdU assays in HPCs treated with MG, LG, HG and HG + DJ-1, respectively. (E) The cell apoptosis was analyzed by Annexin V-FITC/PI apoptosis assay in HPCs treated with MG, LG, HG and HG + DJ-1, respectively. (F) The apoptotic rate of HPCs treated with MG, LG, HG and HG + DJ-1 was analyzed by GraphPad Prism 8 software. (G) Representative Western blot images and quantitative statistical bar graphs of protein expression levels of cleaved caspase 3 and Bcl2 in HPCs treated with LG, HG and HG + DJ-1 were analyzed by Western blotting analysis and Image J software, respectively. MG: mannitol, LG: 5.5mM D-glucose, MG: 15mM D-glucose, HG: 30mM D-glucose, DJ-1: transfected with 1 μg/mL DJ-1-Flag-pcDNA3.1 plasmid. Data are shown as mean ± SD and representative of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) apoptosis detection kit (HY-K1073, MCE, NJ, USA) was carried out to measure HPC apoptosis.

Techniques: Western Blot, Expressing, Software, Quantitative RT-PCR, CCK-8 Assay, Apoptosis Assay, Transfection, Plasmid Preparation

Journal: PLOS One

Article Title: DJ-1 alleviates high glucose-induced podocyte injury via activating ERK1/2 signaling

doi: 10.1371/journal.pone.0346714

Figure Lengend Snippet: (A) Representative Western blot images and quantitative statistical bar graphs of protein expression levels of NF-κB p65 and AP-1 in HPCs treated with LG, HG, HG + DJ-1 and HG + DJ-1 + temuterkib were analyzed by Western blotting analysis and Image J software, respectively. (B) Representative Western blot images and quantitative statistical bar graphs of protein expression levels of cleaved caspase 3 and Bcl2 in HPCs treated with LG, HG, HG + DJ-1 and HG + DJ-1 + temuterkib were analyzed by Western blotting analysis and Image J software, respectively. (C) The cell viability was measured by CCK-8 assays in HPCs treated with LG, HG, HG + DJ-1 and HG + DJ-1 + temuterkib, respectively. (D) The cell viability was analyzed by EdU assays in HPCs treated with LG, HG, HG + DJ-1 and HG + DJ-1 + temuterkib, respectively. (E) The cell apoptosis was analyzed by Annexin V-FITC/PI apoptosis assay in HPCs treated with LG, HG, HG + DJ-1 and HG + DJ-1 + temuterkib, respectively. (F) The apoptotic rate of HPCs treated with LG, HG, HG + DJ-1 and HG + DJ-1 + temuterkib were analyzed by GraphPad Prism 8 software. MG: mannitol, LG: 5.5mM D-glucose, MG: 15mM D-glucose, HG: 30mM D-glucose,temuterkib: 1μM, DJ-1: transfected with 1 μg/mL DJ-1-Flag-pcDNA3.1 plasmid. Data are shown as mean ± SD and representative of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) apoptosis detection kit (HY-K1073, MCE, NJ, USA) was carried out to measure HPC apoptosis.

Techniques: Western Blot, Expressing, Software, CCK-8 Assay, Apoptosis Assay, Transfection, Plasmid Preparation

Journal: Journal of Cardiovascular Pharmacology

Article Title: Integrating Network Pharmacology and In Vitro 3D Cardiac Organoid Model Validation to Investigate the Action Mechanism of Shenfu Xiangshao Decoction on Heart Failure

doi: 10.1097/FJC.0000000000001802

Figure Lengend Snippet: The effects of QU and KA on inflammation, apoptosis, and fibrosis in HF COs. The levels of IL-6 and TNF-α in the supernatant are examined by ELISA kits under the treatment of QU (A, B) or KA (C, D). E, TUNEL staining is applied to detect cardiomyocyte apoptosis after QU or KA treatment. Red fluorescence indicates apoptotic cells, while blue fluorescence marks cell nuclei. Scale bars, 100 μm. The relative fluorescence intensities of QU (G) or KA (H) treatments are quantified. F, The Masson staining of COs sections treated by QU or KA. Collagen fibers are blue, and the cytoplasm of cardiomyocytes is red. Scale bars, 200 μm (upper panel) and 50 μm (lower panel). The relative collagen content of QU (I) or KA (J) treatments is quantified. Values are presented as mean ± SEM (n = 3). * P < 0.05, ** P < 0.01, **** P < 0.0001 versus Control group, # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001 versus Model group.

Article Snippet: The terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay (Fluorescein TUNEL Cell Apoptosis Detection Kit, Servicebio, G1501) was used to assess the apoptosis in COs.

Techniques: Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining, Fluorescence, Control

Journal: Science Advances

Article Title: A conserved glycan motif induces broadly reactive functional antibodies against the zoonotic pathogen Streptococcus suis

doi: 10.1126/sciadv.adz1854

Figure Lengend Snippet: ( A ) Binding of plant lectins to surface structures of capsule-deficient S. suis strains (ΔCPS). SBA binds to N -acetylgalactosamine (GalNAc) and, to a lesser extent, galactose (Gal); RCA 120 binds to both Gal and GalNAc; sWGA has a special affinity to N -acetylglucosamine (GlcNAc). Data from biological triplicates are presented as mean values ± SD. ( B ) Glycosyl composition analysis by GC-MS of TMS (trimethylsilyl) derivatives of methyl glycosides of S. suis RPS from S10 and 861160 released by mild acid hydrolysis after chemical N -acetylation. ( C ) Plant lectin binding to isolated RPS from S. suis S10 and 861160. Data show technical triplicates (mean values ± SD) and are representative for two independent experiments. ( D and E ) Presence and absence of the most abundant glycosyl linkage residues (D) and phosphate (E) of S. suis RPS from S10 and 861160. Glycosyl linkage residues were analyzed by GC-MS of partially methylated alditol acetate derivatives. Phosphate was determined by malachite green assay following hydrolysis with hydrochloric acid and digestion with alkaline phosphatase. Original data is in tables S2 and S3.

Article Snippet: Subsequently, 12.5 μl of bacterial suspension was incubated in a 96-well V-bottom plate with the same volume of pig serum or fluorescein-labeled lectin (Vector Laboratories, FLK-2100 and FL-1021S-5).

Techniques: Binding Assay, Gas Chromatography-Mass Spectrometry, Isolation, Methylation, Malachite Green Assay