Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Design, synthesis, biological evaluation and molecular docking study of 2,4-diarylimidazoles and 2,4-bis(benzyloxy)-5-arylpyrimidines as novel HSP90 N-terminal inhibitors

doi: 10.1080/14756366.2022.2124407

Figure Lengend Snippet: Western blotting assay. (a) (b) Effects of 16l (L: 6 μM; H: 18 μM) and 22k (L: 8 μM; H: 24 μM) on the expression of AKT, ERK, HSP70, and HSP90 in MCF-7 cells, with GAPDH as an internal reference. 17-AAG (15 μM) was used as a positive control. (c) (d) Statistical analysis of western blotting assays of 16l and 22k , respectively. Data are presented as the means ± SDs. * p < 0.05, ** p < 0.01, *** p < 0.001 versus control.

Article Snippet: Primary antibodies against ERK and AKT were purchased from Beyotime (Shanghai, China).

Techniques: Western Blot, Expressing, Positive Control, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Melatonin Protects Intervertebral Disc from Degeneration by Improving Cell Survival and Function via Activation of the ERK1/2 Signaling Pathway

doi: 10.1155/2019/5120275

Figure Lengend Snippet: Melatonin, via the ERK pathway, regulates NPCs. (a–c) Melatonin significantly increased ERK1/2 activity above the concentration of 1 μ m. A slight inhibition of phosphorylated NF- κ B protein expression was detected at 1 μ M. A decrease in phosphorylation level of NF- κ B was also found at high concentrations of melatonin. No obvious change was observed in JNK activity after melatonin treatment. (d) Melatonin increased the proportion of cells in the S-phase and decreased that of cells in the G0/G1 phase. With U0126, the effects on cell cycle distribution were reduced. (e) Similarly, the expression of the cell cycle-related proteins was reduced in NPCs treated with melatonin and U0126. (f) Quantified analysis of cell cycle-related protein Western blot result. (g, h) The inhibition of apoptosis by melatonin was also reversed upon addition of U0126. (i) Quantified analysis of apoptosis-related protein Western blot result. (j) Western blot analysis also showed that, after addition of U0126, U0126 had a significant weakened role on melatonin in the downregulation of collagen X expression and there is no more positive effect from melatonin on aggrecan and collagen II expression in the NPCs. (k) Quantified analysis of cell matrix protein Western blot result ( ∗ p < 0.05).

Article Snippet: For immunohistochemistry detection, sections were incubated with primary antibodies specific to ERK (EterLife, UK), collagen II (Novus Biologicals, CO, USA), and collagen X (Biosynthesis Biotech, China).

Techniques: Activity Assay, Concentration Assay, Inhibition, Expressing, Phospho-proteomics, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Melatonin Protects Intervertebral Disc from Degeneration by Improving Cell Survival and Function via Activation of the ERK1/2 Signaling Pathway

doi: 10.1155/2019/5120275

Figure Lengend Snippet: (a) Magnetic resonance imaging findings after melatonin injection on rabbit intervertebral discs. T2 signal intensity was stronger for the melatonin-injected discs than for the saline discs, including the melatonin plus U01256 group. (b) Thompson score showed the significant radiological improvement on the 8th week brought by melatonin ( ∗ p < 0.05). (c) Histological analysis on intervertebral discs by HE staining. More chondrocytes with intact annulus fibrosus were observed in the melatonin-treated rats. However, reduced number of chondrocytes and twisted or damaged annulus fibrosus presented in the two control groups. ERK 1/2 immunohistochemical staining indicates that ERK 1/2 expression was more apparent in the melatonin group. Quantification of immunohistochemical staining showed that the melatonin group had significantly higher mean optical density than the inhibitor-treated and control groups. Collagen II and X immunohistochemical staining indicates that collagen II was detected in the melatonin-treated group, but little in the two control groups. By contrast, the staining for collagen X was weaker in the melatonin-treated group. Quantification of collagen II immunohistochemical staining showed that the melatonin-treated discs had significantly higher mean optical density than the control groups and the expression of collagen X was significantly lower. (d) Quantified analysis of immunohistochemical staining showed the significant histological improvement brought by melatonin via the ERK pathway ( ∗ p < 0.05).

Article Snippet: For immunohistochemistry detection, sections were incubated with primary antibodies specific to ERK (EterLife, UK), collagen II (Novus Biologicals, CO, USA), and collagen X (Biosynthesis Biotech, China).

Techniques: Magnetic Resonance Imaging, Injection, Saline, Staining, Control, Immunohistochemical staining, Expressing