Journal: Journal of orthopaedic surgery (Hong Kong)

Article Title: Naringenin alleviates bone cancer pain via NF-κB/uPA/PAR2 pathway in mice.

doi: 10.1177/10225536241266671

Figure Lengend Snippet: Figure 4. Naringenin inhibited the level of NF-κB/uPA pathway in BCP The level of uPA in serum and bone tissue and the levels of NF-κB pathway-related proteins containing p-IKKβ, IKKβ, p-p65 and p65 were determined in this part. (a), the level of uPA in serum measured by ELISA. (b), the level of uPA in bone tissue measured by ELISA. (c), the levels of p-IKKβ, IKKβ, p-p65 and p65 using western blot. (d), the levels of uPA and p-p65 using immunocytochemistry. The data are presented as the mean ± standard deviation. n = 8. *p < .05, **p < .01 and ***p < .001.

Article Snippet: Then, membrane was blocked by blocking buffer (Beyotime, China) at 4°C for 4 h, followed by the incubationwith the primary antibodies of TNFα (ab183218, 1:1000), TNFR1 (Abcam, ab223352, 1:1000), TNFRSF1A Associated Via Death Domain (TRADD) (Abcam, ab110644, 1:1000), RIP (Abcam, ab202985, 1:1000), TNF Receptor Associated Factor 2 (TRAF2) (Abcam, ab244317, 0.4 μg/mL), p-Inhibitor of Nuclear Factor Kappa B Kinase Subunit Beta (p-IKKβ) (Cell Signaling, #2697, 1:1000), IKKβ (Cell Signaling, #8943, 1:1000), p-p65 (Cell Signaling, #3033, 1:1000), p65 (Cell Signaling, #8242, 1:1000), PAR2 (Abcam, ab180953, 1:10,000), PKC-γ (Abcam, ab71558, 1: 2000), PKA (Cell Signaling, #4782, 1:1000) and TRPV1 (Abcam, ab6166, 1:1000) for 12 h at 4°C.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Immunocytochemistry, Standard Deviation

Journal: Plants

Article Title: A Comprehensive Review on Cannabis sativa Ethnobotany, Phytochemistry, Molecular Docking and Biological Activities

doi: 10.3390/plants12061245

Figure Lengend Snippet: Molecular docking of Cannabis sativa L. compounds.

Article Snippet: Inhibitor of nuclear factor kappa-b kinase subunit β (IKKbeta) , 3BRT (p) , Inhibitor kappa kinase , Anti-inflammatory , CBD , Cannabinoids , AutoDock-tools , −7.99 , , [ ] .

Techniques: Activity Assay

Journal: Journal of Clinical and Translational Hepatology

Article Title: Retrorsine Cooperates with Gut Microbiota to Promote Hepatic Sinusoidal Obstruction Syndrome by Disrupting the Gut Barrier

doi: 10.14218/JCTH.2021.00398

Figure Lengend Snippet: (A) Representative ileal hematoxylin and eosin staining ( n =6 per group). Scale bar, 100 µm. (B, C) mRNA expression of innate immune components ( Cd14 , Tlr4 , Tlr9 ) and inflammatory cytokines ( Il6 , ccl2 , Ccl4 , Cxcl2 , and Icam1 ) in the distal ileum of mice treated with RTS for 24 h with or without ABX pretreatment ( n =6 per group). (D) Protein expression of IKKβ, p-IKKβ, p65, and p-p65 in the distal ileum. GAPDH was the loading control ( n= 3). (E) mRNA expression of proinflammatory factors ( Il6 , Tnfα , Ccl2 , Ccl4 and Cxcl2 ) in HCT116 cells ( n= 3. (F) Protein expression of IKKβ, p-IKKβ, p65, and p-p65 in HCT116 cells. GAPDH was the loading control ( n= 3. (G) Relative expression of IKKβ, p-IKKβ, p65, and p-p65 in HCT116 cells ( n= 3). Veh, control group treated with vehicle; LPS group, HCT116 cells stimulated with LPS (5 µg/mL) for 24 h; RTS group, HCT116 cells stimulated with RTS (600 µM) for 24 h; RTS + LPS group included HCT116 cells costimulated with RTS (600 µM) and LPS (5 µg/mL) for 24 h. Data are means±standard error of the mean. * p< 0.05, ** p< 0.01; analysis of variance. ABX, nonabsorbable antibiotics; LPS, lipopolysaccharides; ns, not significant; RTS, retrorsine; Veh, vehicle.

Article Snippet: After blocking in 5% BSA/TBST for 1 h at room temperature, membranes were incubated with primary antibodies overnight at 4°C: ZO-1, 1:500, (ab96587; Abcam); occludin, 1:1,000 (331594; ThermoFisher Scientific); PV1, 1:500 (ab27853; Abcam); inhibitor of nuclear-factor kappa-B kinase subunit beta (IKKβ), 1:1,000 (8943; Cell Signaling Technology, Danvers, MA, USA); p-IKKβ, 1:1,000 (2697; Cell Signaling Technology); p65, 1:1,000 (3022; Cell Signaling Technology); p-p65, 1:1,000 (3031; Cell Signaling Technology); GAPDH, 1:3,000 (ANT012; Antgene); Actin, 1:2,000 (ANT010; Antgene), followed by incubation with corresponding secondary antibody (1:3,000; Antgene) for 1 h at room temperature.

Techniques: Staining, Expressing, Control

Journal: Journal of Clinical and Translational Hepatology

Article Title: Retrorsine Cooperates with Gut Microbiota to Promote Hepatic Sinusoidal Obstruction Syndrome by Disrupting the Gut Barrier

doi: 10.14218/JCTH.2021.00398

Figure Lengend Snippet: (A) Serum IL6 levels in mice ( n= 8 per group). (B) Serum TNFα levels in mice ( n= 8 per group). (C) Western blots and (D) relative expression of IKKβ, p-IKKβ, P65. and p-P65 protein in the liver. GAPDH was the loading control ( n= 3 per group). Data are means±standard error of the mean. ns: no significance, * p< 0.05, ** p< 0.01; analysis of variance. ABX, nonabsorbable antibiotics; RTS, retrorsine; Veh, vehicle.

Article Snippet: After blocking in 5% BSA/TBST for 1 h at room temperature, membranes were incubated with primary antibodies overnight at 4°C: ZO-1, 1:500, (ab96587; Abcam); occludin, 1:1,000 (331594; ThermoFisher Scientific); PV1, 1:500 (ab27853; Abcam); inhibitor of nuclear-factor kappa-B kinase subunit beta (IKKβ), 1:1,000 (8943; Cell Signaling Technology, Danvers, MA, USA); p-IKKβ, 1:1,000 (2697; Cell Signaling Technology); p65, 1:1,000 (3022; Cell Signaling Technology); p-p65, 1:1,000 (3031; Cell Signaling Technology); GAPDH, 1:3,000 (ANT012; Antgene); Actin, 1:2,000 (ANT010; Antgene), followed by incubation with corresponding secondary antibody (1:3,000; Antgene) for 1 h at room temperature.

Techniques: Western Blot, Expressing, Control