Journal: iScience

Article Title: Disease phenotypic screening in neuron-glia cocultures identifies blockers of inflammatory neurodegeneration

doi: 10.1016/j.isci.2024.109454

Figure Lengend Snippet:

Article Snippet: BAY61-3606 (hydrochloride) , Cayman Chemical , 11423.

Techniques: Recombinant, Software

Journal: Frontiers in Pharmacology

Article Title: Luteolin inhibits GPVI-mediated platelet activation, oxidative stress, and thrombosis

doi: 10.3389/fphar.2023.1255069

Figure Lengend Snippet: Effect of luteolin on GPVI-mediated oxidative stress in human platelets. The reactive oxygen species (ROS) assay probes DCFH-DA (5 μM), 2 μM Amplex Red, and 2 μM DHE were used to pre-stained human washed platelets (WPs). The WPs were incubated with different concentrations of luteolin, DMSO, and Syk-specific inhibitor (BAY61-3606) for 10 min at 37°C. (A) Collagen (10 μg/mL) and GPVI-specific activator convulxin (5 ng/mL) activation of human platelets. Human platelet ROS release was monitored in real-time at ex/em = 480/530 nm. RFU: relative fluorescence value. (B,C) Flow cytometry detection of platelet superoxide anion and H 2 O 2 release. (D) Effect of luteolin and VAS2870 (10 μM) on collagen (10 μg/mL) and convulxin (5 ng/mL) -induced NOXs activity. (E–G) Platelet lysate supernatant MDA, SOD, and GPx activities were measured. (H) Effect of luteolin (25 μM) and BAY61-3606 (63 nM) on collagen- (10 μg/mL) and convulxin (5 ng/mL)-induced platelet ROS release. Absorbance values were detected at 450 nm after the reaction of platelet membrane protein lysate with NADPH (0.5 mM) and lucigenin (10 μM). Compared with the BAY61-3603 group, # p < 0.05, ### p < 0.001. Data were analyzed with one-way ANOVA followed by Dunnett’s test. Values are expressed as mean ± standard error, n ≥ 3. Compared with the Vehicle (DMSO) group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The platelets were then incubated with luteolin 2.5, 10, or 25 μM, BAY61-3606 (63 nM, Cat No.HY-76474; MCE), BAY61-3606+luteolin 25 μM, or an equal volume of DMSO at 37°C for 10 min.

Techniques: ROS Assay, Staining, Incubation, Activation Assay, Fluorescence, Flow Cytometry, Activity Assay, Membrane

Journal: Frontiers in Pharmacology

Article Title: Luteolin inhibits GPVI-mediated platelet activation, oxidative stress, and thrombosis

doi: 10.3389/fphar.2023.1255069

Figure Lengend Snippet: Effect of luteolin on GPVI-mediated oxidative stress in human platelets. The reactive oxygen species (ROS) assay probes DCFH-DA (5 μM), 2 μM Amplex Red, and 2 μM DHE were used to pre-stained human washed platelets (WPs). The WPs were incubated with different concentrations of luteolin, DMSO, and Syk-specific inhibitor (BAY61-3606) for 10 min at 37°C. (A) Collagen (10 μg/mL) and GPVI-specific activator convulxin (5 ng/mL) activation of human platelets. Human platelet ROS release was monitored in real-time at ex/em = 480/530 nm. RFU: relative fluorescence value. (B,C) Flow cytometry detection of platelet superoxide anion and H 2 O 2 release. (D) Effect of luteolin and VAS2870 (10 μM) on collagen (10 μg/mL) and convulxin (5 ng/mL) -induced NOXs activity. (E–G) Platelet lysate supernatant MDA, SOD, and GPx activities were measured. (H) Effect of luteolin (25 μM) and BAY61-3606 (63 nM) on collagen- (10 μg/mL) and convulxin (5 ng/mL)-induced platelet ROS release. Absorbance values were detected at 450 nm after the reaction of platelet membrane protein lysate with NADPH (0.5 mM) and lucigenin (10 μM). Compared with the BAY61-3603 group, # p < 0.05, ### p < 0.001. Data were analyzed with one-way ANOVA followed by Dunnett’s test. Values are expressed as mean ± standard error, n ≥ 3. Compared with the Vehicle (DMSO) group, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The platelets were then incubated with luteolin 2.5, 10, or 25 μM, BAY61-3606 (63 nM, Cat No.HY-76474; MCE), BAY61-3606+luteolin 25 μM, or an equal volume of DMSO at 37°C for 10 min.

Techniques: ROS Assay, Staining, Incubation, Activation Assay, Fluorescence, Flow Cytometry, Activity Assay, Membrane