Journal: Drug Design, Development and Therapy

Article Title: Obacunone Promotes Functional Recovery After Spinal Cord Injury by Attenuating Neuroinflammation by Targeting the TLR4/MyD88/p38 MAPK Pathway

doi: 10.2147/DDDT.S577707

Figure Lengend Snippet: Oba inhibits the TLR4/MyD88/p38 MAPK signaling pathway in both in vivo and in vitro experiments. ( A – D ) Molecular docking analysis of Oba with TLR4. ( E – H ) Molecular docking analysis of Oba with MyD88. ( I – L ) Molecular docking analysis of Oba with p38. For each protein, the analysis is presented as: an overall docking pose with the binding region outlined by a red dashed box ( A, E, I ); a close-up view of the binding region ( B, F, J ); a surface representation ( C, G, K ); and a two-dimensional interaction diagram ( D, H, L ). ( M ) Representative Western blot image of TLR4, MyD88, p-p38 and p38 protein in BV-2 cells. The plus and minus signs (+, -) denote the presence or absence of the following reagents in culture: LPS, and 50 μM Oba. ( N ) Quantitative analysis of TLR4 and MyD88 protein levels and p38 phosphorylation in BV-2 cells (n = 5). ( O ) Western blot analysis of TLR4, MyD88, p-p38, and p38 in spinal cord tissues from Sham, SCI, and SCI + Oba (20 mg/kg) groups at 3 days post-SCI. ( P ) Quantitative analysis of TLR4 and MyD88 protein levels and p38 phosphorylation in spinal cord tissues (n = 5). The values are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001; NS, not significant.

Article Snippet: To investigate the role of the p38 MAPK pathway in the anti-inflammatory effects of Oba, BV-2 cells were pre-treated for 3 hours with 50 μM Oba, 10 μM SB203580 (a p38 MAPK inhibitor; HY-10256, MCE, China), or a combination of both, prior to stimulation with LPS for 24 hours.

Techniques: In Vivo, In Vitro, Binding Assay, Western Blot, Phospho-proteomics

Journal: Drug Design, Development and Therapy

Article Title: Obacunone Promotes Functional Recovery After Spinal Cord Injury by Attenuating Neuroinflammation by Targeting the TLR4/MyD88/p38 MAPK Pathway

doi: 10.2147/DDDT.S577707

Figure Lengend Snippet: Oba attenuates inflammation by targeting the TLR4/MyD88/p38 MAPK pathway in BV-2 microglial cells. ( A ) Representative Western blot of TLR4 expression in BV-2 cells transfected with a TLR4 overexpression plasmid (pcDNA3.1-TLR4) or an empty vector. The plus and minus signs (+, -) denote the presence or absence of the following reagents in culture: empty plasmid (Oe-NC), and TLR4 overexpression plasmid (Oe-TLR4). ( B ) Quantitative analysis of TLR4 protein levels (n = 5). ( C ) Western blot images of iNOS, TLR4, TNFα and IL-1β protein expression in BV-2 cells. The plus and minus signs (+, -) denote the presence or absence of the following treatments in culture: Oe-NC, Oe-TLR4, LPS, and Oba. ( D – G ) Quantitative analysis of iNOS, TLR4, TNFα and IL-1β protein levels (n = 5). ( H ) Representative Western blot images of iNOS, TNFα, IL-1β, p-p38 and p38 in BV-2 cells. The plus and minus signs (+, -) denote the presence or absence of the following reagents in culture: LPS, Oba, and SB203580. ( I – L ) Quantitative analysis of iNOS, TNFα, IL-1β protein levels and p38 phosphorylation (n = 5). The values are presented as mean ± SD. *p<0.05, **p<0.01, ***p<0.001; NS, not significant.

Article Snippet: To investigate the role of the p38 MAPK pathway in the anti-inflammatory effects of Oba, BV-2 cells were pre-treated for 3 hours with 50 μM Oba, 10 μM SB203580 (a p38 MAPK inhibitor; HY-10256, MCE, China), or a combination of both, prior to stimulation with LPS for 24 hours.

Techniques: Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation, Phospho-proteomics

Journal: Journal of Musculoskeletal & Neuronal Interactions

Article Title: HOXA7 Impairs Osteogenic Differentiation via p38/JNK Signaling: Implications for Osteoporosis

doi: 10.22540/JMNI-26-134

Figure Lengend Snippet: HOXA7 regulates the p38 MAPK/JNK pathway in hBMSCs and MC3T3-E1 cells. (A–B) Phosphorylated and total p38 MAPK and JNK (p-p38/p38, p-JNK/JNK) in hBMSCs and MC3T3-E1 cells with the indicated transfections were evaluated by Western blotting. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: To assess the p38 MAPK/JNK pathway, the JNK inhibitor SP600125 (cat. no. 8177; Cell Signaling Technology) and the p38 inhibitor SB203580 (cat. no. 5633; Cell Signaling Technology) were first dissolved in DMSO and then added to the MC3T3-E1 cells at a desired concentration of 20 μM; the cells were then incubated at 37 °C in 5% CO 2 for 3 days.

Techniques: Transfection, Western Blot

Journal: Journal of Musculoskeletal & Neuronal Interactions

Article Title: HOXA7 Impairs Osteogenic Differentiation via p38/JNK Signaling: Implications for Osteoporosis

doi: 10.22540/JMNI-26-134

Figure Lengend Snippet: Effects of SP600125 and SB203580 on proliferation, apoptosis, and autophagy in MC3T3-E1 cells after HOXA7 knockdown. (A) Western blotting of p-JNK/JNK and p-p38/p38 in MC3T3-E1 cells under the indicated conditions. (B) CCK-8 assay assessing cell viability. (C) Flow cytometry analysis of apoptosis. (D) Western blotting of ATG12, p62, and LC3-II/I. Significance vs. si-NC: **P < 0.01, ***P < 0.001, ****P < 0.0001. Significance vs. si-HOXA7: ##P < 0.01, ###P < 0.001, ####P < 0.0001.

Article Snippet: To assess the p38 MAPK/JNK pathway, the JNK inhibitor SP600125 (cat. no. 8177; Cell Signaling Technology) and the p38 inhibitor SB203580 (cat. no. 5633; Cell Signaling Technology) were first dissolved in DMSO and then added to the MC3T3-E1 cells at a desired concentration of 20 μM; the cells were then incubated at 37 °C in 5% CO 2 for 3 days.

Techniques: Knockdown, Western Blot, CCK-8 Assay, Flow Cytometry