Journal: International Journal of Molecular Sciences

Article Title: Betula alba Bark Extract and Empetrum nigrum Fruit Juice, a Natural Alternative to Niacinamide for Skin Barrier Benefits

doi: 10.3390/ijms232012507

Figure Lengend Snippet: Summary of IC50 of the two extracts BA (circle) and EN (triangle) in ( a ) KLK5 ( n = 3), ( b ) hyaluronidase ( n = 3), ( c ) DPPH ( n = 3) and ( d ) COX2 ( n = 4) inhibition assays. Plotted values are displayed as mean (±SD).

Article Snippet: Thirty microliter of Tris-HCl buffer (0.1 M, pH 8), 10 µL hematin (Sigma Aldrich, H3281), 10 µL of extract (1% final concentration of raw material) and 10 µL enzyme COX2 (BPS Bioscience, San Diego, CA, USA, 71111) 20 ng/µL were introduced into a 96-well plate and incubated 15 min at room temperature.

Techniques: Inhibition

Journal: Journal of Cellular and Molecular Medicine

Article Title: Anti‐inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma

doi: 10.1111/jcmm.18356

Figure Lengend Snippet: TVE specifically suppresses NLRP3 inflammasome. (A) Schematic outline of the NLRP3 inflammasome activation experiments in vitro. (B–C) PMA (500 nM)‐differentiated THP‐1 cells (B) and J774A.1 cells (C) were primed by LPS (100 ng/mL) for 3 h, before were treated with TVE for 2 h. Cell viability was analysed by EZ‐Cytox. (D–E) LPS‐primed THP‐1 cells were treated with TVE for 2 h and then stimulated for 1 h with nigericin (10 μM) (D) and ATP (5 mM) (E) with or without MCC950. (F–G) LPS‐primed J774A.1 cells were treated with TVE for 2 h and then stimulated for 30 min with nigericin (10 μM) (F), ATP (5 mM) (G) with or without MCC950. This data represents the investigation of the protein expression level of IL‐1β in supernatant, confirmed by immunoblot, using densitometry to assess intensity. (H‐J) IL‐1β (p17) and in the supernatants (Sup) and soluble lysates (Lys) were analysed by immunoblot. LPS‐primed J774A.1 cells were treated with TVE for 2 h and then stimulated for 3 h with MSU (100 μg/mL) (H) or dsDNA (2 μg/mL) (I) and flagellin (1.25 μg/mL) (J) by using Lipofectamine 3000™. One representative result of three independent experiments is shown. Values shown are reported as the means of technical triplicates ± SEM. One‐way ANOVA, Bonferroni post‐hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ATP, Adenosine triphosphate; IL‐1β, Interleukin‐1beta; LPS, Lipopolysaccharide; MCC, MCC950; Nig, Nigericin; n.s., not significant; TVE, Trichospira verticillata (L.) S.F. Blake Extract.

Article Snippet: The purified mouse NLRP3 (BPS bioscience, 100189) (0.139 mg/mL) was incubated with TVE (50 μg/mL) or DMSO at 37°C for 30 min in the reaction buffer (100 mM Tris pH 7.8, 2.8 mM EDTA, 100 mM MgCl₂, 15 mM KCl, 655 mM NaCl).

Techniques: Activation Assay, In Vitro, Expressing, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Anti‐inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma

doi: 10.1111/jcmm.18356

Figure Lengend Snippet: TVE suppresses NLRP3 inflammasome activation regardless of K + efflux, ROS and ATPase activity. (A) LPS‐primed J774A.1 cells were treated with TVE for 2 h and then stimulated with IMQ (200 μM) for 1 h. (B) LPS‐primed J774A.1 cells were treated with or without TVE or NAC for 2 h and then stimulated with ATP (5 mM) for 25 min. ROS levels are detected by a microplate reader using a DCFDA solution (20 μM). (C) The amount of NLRP3‐mediated ATP converted into ADP with TVE was determined by luminescence using the ADP‐Glo Assay. (D) and (E) The NLRP3‐Myc transfected‐HEK 293FT cells were treated with or without TVE for 2 h. The NLRP3‐NEK7 interaction was analysed by immunoprecipitation and immunoblot. One representative result of three independent experiments is shown. Values are shown reported as the means of technical triplicates ± SEM. One‐way ANOVA, Bonferroni post‐hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. LPS, Lipopolysaccharide; n.s., not significant; NAC, N‐acetyl‐l‐cysteine; ROS, reactive oxygen species; TVE, Trichospira verticillata (L.) S.F. Blake Extract.

Article Snippet: The purified mouse NLRP3 (BPS bioscience, 100189) (0.139 mg/mL) was incubated with TVE (50 μg/mL) or DMSO at 37°C for 30 min in the reaction buffer (100 mM Tris pH 7.8, 2.8 mM EDTA, 100 mM MgCl₂, 15 mM KCl, 655 mM NaCl).

Techniques: Activation Assay, Activity Assay, Glo Assay, Transfection, Immunoprecipitation, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Anti‐inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma

doi: 10.1111/jcmm.18356

Figure Lengend Snippet: TVE blocks NLRP3 inflammasome assembly by hindering ASC oligomerization. (A) One representative ASC speck (indicated by arrow) images of at least 10 images are shown. LPS‐primed J774A.1 cells were treated with TVE for 2 h and then stimulated with ATP (5 mM) for 30 min. (B) LPS‐primed THP‐1 cells were treated with TVE for 2 h and then stimulated with nigericin (10 μM) for 30 min. ASC oligomerization in the cross‐linked cytosolic pellet was analysed by immunoblot. Scale bar, 20 μm. One representative result of three independent experiments is shown. Values are shown reported as the means of 10 replicates ± SEM. One‐way ANOVA, Bonferroni post‐hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ASC, apoptosis‐associated speck‐like protein; LPS, Lipopolysaccharide; n.s., not significant; TVE, Trichospira verticillata (L.) S.F. Blake Extract.

Article Snippet: The purified mouse NLRP3 (BPS bioscience, 100189) (0.139 mg/mL) was incubated with TVE (50 μg/mL) or DMSO at 37°C for 30 min in the reaction buffer (100 mM Tris pH 7.8, 2.8 mM EDTA, 100 mM MgCl₂, 15 mM KCl, 655 mM NaCl).

Techniques: Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Anti‐inflammatory effect of Trichospira verticillata via suppression of the NLRP3 inflammasome in neutrophilic asthma

doi: 10.1111/jcmm.18356

Figure Lengend Snippet: TVE inhibits NLRP3 inflammasome in lung epithelial cells. (A–B) A549 cells were treated with TVE for 2 h (A), or overnight (B). Cell viability was analysed by EZ‐CYTOX. (C–D) A549 cells and PMA (500 nM)‐differentiated THP‐1 cells were co‐cultured and primed by LPS (100 ng/mL) for 3 h, before they were treated with TVE for 2 h and stimulated for 30 min with nigericin (10 μM) (C), and ATP (5 mM) (D). The level of IL‐1β was evaluated by ELISA and the lysate was analysed by immunoblotting. One representative result of three independent experiments is shown. Values are shown reported as the means of technical triplicates ± SEM. One‐way ANOVA, Bonferroni post‐hoc test; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n.s. not significant.

Article Snippet: The purified mouse NLRP3 (BPS bioscience, 100189) (0.139 mg/mL) was incubated with TVE (50 μg/mL) or DMSO at 37°C for 30 min in the reaction buffer (100 mM Tris pH 7.8, 2.8 mM EDTA, 100 mM MgCl₂, 15 mM KCl, 655 mM NaCl).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot