Journal: Dentistry Journal

Article Title: In Vitro Assessment of Osteogenic Modulation and Molecular Responses Induced by Contemporary Endodontic Sealers in MC3T3-E1 Pre-Osteoblasts

doi: 10.3390/dj14030160

Figure Lengend Snippet: Effects of ZOE and BC sealers on MC3T3-E1 cell viability and MAPK signaling. ( A ) Western blot analysis of phosphorylated and total c-Jun, p38, and Erk in cells cultured with ZOE or BC for 1 week. Gapdh (loading control). Phospho-c-Jun, p38, and Erk band intensities were normalized to their respective total protein levels. ( B ) Representative images of MC3T3-E1 cells cultured in the presence of ZOE or BC for 4 and 5 days. Scale bars: 100 μm (white), 50 μm (black). ( C , D ) Quantification of cell numbers around sealer spots at day 4 ( C ) and day 5 ( D ). The number of cells in five random fields was counted per experimental condition under light microscopy. Data are presented as mean ± SD, and the experiments were repeated three times. Statistical significance was assessed by Student’s t -test (** p < 0.001). ( E , F ) Percentages of viable ( E ) and non-viable ( F ) cells on day 5 determined by trypan blue exclusion. Data are presented as mean ± SD from triplicate assays, and the experiments were repeated three times. Statistical significance was assessed by Student’s t -test (** p < 0.001).

Article Snippet: Primary antibodies purchased from Cell Signaling Technology were phospho (p)-Erk1/2 (1:2000), Erk1/2 (1:2000), p-p38 (1:2000), p38 (1:2000), p-c-Jun/Ser73 (1:2000), c-Jun (1:1000), Runx2 (1:2000), Gapdh (1:5000), and from Santa-Cruz Biotechnology was Osx (1:1000).

Techniques: Western Blot, Cell Culture, Control, Light Microscopy

Journal: Dentistry Journal

Article Title: In Vitro Assessment of Osteogenic Modulation and Molecular Responses Induced by Contemporary Endodontic Sealers in MC3T3-E1 Pre-Osteoblasts

doi: 10.3390/dj14030160

Figure Lengend Snippet: Effects of MAPK inhibition on BC-induced osteogenic differentiation in MC3T3-E1 cells. ( A , B ) qRT-PCR analysis of Runx2 ( A ) and Sp7 ( Osx ) ( B ) expression after 1 week of culture in OI medium under control conditions (no sealer), BC, or BC combined with MAPK inhibitors [U0126 (Erk), SB203580 (p38), SP600125 (Jnk)]. Expression levels were normalized to Gapdh . Data are presented as mean ± SD from triplicate assays, and the experiments were repeated three times. Statistical significance was assessed by one-way ANOVA with Tukey’s post hoc test (* p < 0.01). ( C ) Western blot analysis of Runx2, Sp7 (Osx), p-c-Jun, c-Jun, p-p38, p38, p-Erk, and Erk under the same conditions as ( A ). Gapdh (loading control). ( D ) Representative of ARS staining images of cells cultured with control (no sealer), BC, or BC plus MAPK inhibitors for 2 weeks with OIM. Scale bars = 100 μm. ( E ) Quantification of ARS staining intensity using ImageJ. Data are presented as mean ± SD from 5 fields, and the experiments were repeated three times. Statistical significance was assessed by one-way ANOVA with Tukey’s post hoc test (*** p < 0.0001). ( F ) Proposed schematic model, based on the present findings, illustrating that the BC sealer regulates MAPK signaling pathways (Jnk, p38, Erk) to induce osteogenic differentiation by modulating the expression of Runx2 and Sp7 ( Osx ) in pre-osteoblasts.

Article Snippet: Primary antibodies purchased from Cell Signaling Technology were phospho (p)-Erk1/2 (1:2000), Erk1/2 (1:2000), p-p38 (1:2000), p38 (1:2000), p-c-Jun/Ser73 (1:2000), c-Jun (1:1000), Runx2 (1:2000), Gapdh (1:5000), and from Santa-Cruz Biotechnology was Osx (1:1000).

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Control, Western Blot, Staining, Cell Culture, Protein-Protein interactions

Journal: Frontiers in Molecular Biosciences

Article Title: Contribution of Increased Expression of Yin Yang 2 to Development of Cardiomyopathy

doi: 10.3389/fmolb.2020.00035

Figure Lengend Snippet: Altered JNK signaling pathways in double transgenic (dTg) mouse hearts. Western blot analysis was performed by using heart lysates from control and dTg mice. GAPDH was used as a loading control. (A) p38 activity was not significantly altered between age-matched control and dTg hearts. (B) Quantification of the data in (A) . (C) JNK activity was significantly higher in dTg hearts than in control hearts. (D) Quantification of the data shown in (C) . ∗ p < 0.05 vs. Cont. n = 3 per group.Cont, control; dTg, pCAG-YY2-Tg+/-MHC-Cre+.

Article Snippet: Antibodies against p38 (Cat# AF8691, 1:1000) and p38 phosphorylated on Thr180 and Tyr182 (p-p38, Cat# NB500-138, 1:1000) were from Novus Biologicals (Centennial, CO, United States).

Techniques: Protein-Protein interactions, Transgenic Assay, Western Blot, Control, Activity Assay