Journal: The Journal of International Medical Research

Article Title: Hypomethylated leptin receptor reduces cerebral ischaemia–reperfusion injury by activating the JAK2/STAT3 signalling pathway

doi: 10.1177/03000605241261912

Figure Lengend Snippet: Primer sequences for real-time quantitative polymerase chain reaction analyses.

Article Snippet: The membranes were incubated in blocking solution consisting of Tris-buffered saline-Tween 20 (TBST; pH 7.5; 20 mmol/l Tris–HCl, 150 mmol/l sodium chloride, 0.1% Tween-20) containing 5% nonfat milk at room temperature for 1 h. The membranes were then incubated with primary rabbit antirat antibodies for JAK2, STAT3, p-JAK3, p-STAT3, IL-6, IL-10, caspase 3, BCL-2 and GAPDH (dilution 1:1000; Beyotime Biotechnology) at 4°C overnight.

Techniques: Real-time Polymerase Chain Reaction

Journal: The Journal of International Medical Research

Article Title: Hypomethylated leptin receptor reduces cerebral ischaemia–reperfusion injury by activating the JAK2/STAT3 signalling pathway

doi: 10.1177/03000605241261912

Figure Lengend Snippet: (a) PC12 neuronal cell viability under different leptin concentrations (50–300 ng/ml) for 30 min after the OGD for 1 h and then the reperfusion of 24 h; (b) PC12 neuronal cell viability following different reperfusion times (1 h, 6 h, 12 h and 24 h) subsequent to pretreatment with different leptin concentrations (50–300 ng/ml) and (c, d) mRNA levels of the inflammatory factor interleukin (IL)-6 and the anti-inflammatory factor IL-10 relative to levels of GAPDH. Data were analysed using one-way analysis of variance; * P < 0.05; *** P < 0.001; compared with the control group. GAPDH, glyceraldehyde 3-phosphate dehydrogenase. The colour version of this figure is available at: http://imr.sagepub.com .

Article Snippet: The membranes were incubated in blocking solution consisting of Tris-buffered saline-Tween 20 (TBST; pH 7.5; 20 mmol/l Tris–HCl, 150 mmol/l sodium chloride, 0.1% Tween-20) containing 5% nonfat milk at room temperature for 1 h. The membranes were then incubated with primary rabbit antirat antibodies for JAK2, STAT3, p-JAK3, p-STAT3, IL-6, IL-10, caspase 3, BCL-2 and GAPDH (dilution 1:1000; Beyotime Biotechnology) at 4°C overnight.

Techniques: Control

Journal: The Journal of International Medical Research

Article Title: Hypomethylated leptin receptor reduces cerebral ischaemia–reperfusion injury by activating the JAK2/STAT3 signalling pathway

doi: 10.1177/03000605241261912

Figure Lengend Snippet: The levels of interleukin (IL)-10, IL-6, tumour necrosis factor (TNF)-α, JAK2 and STAT3 mRNA relative to levels of GAPDH in the brain tissues (a–e). The data were analysed using one-way analysis of variance; ### P < 0.001 compared with the SHAM group; ** P < 0.01, *** P < 0.001, compared with the MCAO group. JAK2, Janus kinase 2; STAT3, signal transducer and activator of transcription 3; reverse transcription-quantitative polymerase chain reaction; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; SHAM, sham-operated group; MCAO, middle cerebral artery occlusion group; MCAO + LP, MCAO group pretreated with leptin. The colour version of this figure is available at: http://imr.sagepub.com .

Article Snippet: The membranes were incubated in blocking solution consisting of Tris-buffered saline-Tween 20 (TBST; pH 7.5; 20 mmol/l Tris–HCl, 150 mmol/l sodium chloride, 0.1% Tween-20) containing 5% nonfat milk at room temperature for 1 h. The membranes were then incubated with primary rabbit antirat antibodies for JAK2, STAT3, p-JAK3, p-STAT3, IL-6, IL-10, caspase 3, BCL-2 and GAPDH (dilution 1:1000; Beyotime Biotechnology) at 4°C overnight.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: The Journal of International Medical Research

Article Title: Hypomethylated leptin receptor reduces cerebral ischaemia–reperfusion injury by activating the JAK2/STAT3 signalling pathway

doi: 10.1177/03000605241261912

Figure Lengend Snippet: Western blot analysis showing the protein levels of phosphorylated-STAT3 (p-STAT3) (a), phosphorylated-JAK2 (p-JAK2) (b), anti-apoptotic protein BCL-2 (c) and apoptotic protein caspase 3 (d) relative to levels of GAPDH. Western blot analysis showing the levels of P-JAK2, P-STAT3, caspase 3, BCL-2 and GAPDH in the SHAM, MCAO and MCAO + LP groups. GAPDH was used as a loading control (e). The data were analysed using one-way analysis of variance; # P < 0.05, ## P < 0.01, compared with the SHAM group; * P < 0.05, ** P < 0.01, compared with the MCAO group; STAT3, signal transducer and activator of transcription 3; JAK2, Janus kinase 2; BCL-2, B-cell lymphoma 2; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; SHAM, sham-operated group; MCAO, middle cerebral artery occlusion group; MCAO + LP, MCAO group pretreated with leptin. The colour version of this figure is available at: http://imr.sagepub.com .

Article Snippet: The membranes were incubated in blocking solution consisting of Tris-buffered saline-Tween 20 (TBST; pH 7.5; 20 mmol/l Tris–HCl, 150 mmol/l sodium chloride, 0.1% Tween-20) containing 5% nonfat milk at room temperature for 1 h. The membranes were then incubated with primary rabbit antirat antibodies for JAK2, STAT3, p-JAK3, p-STAT3, IL-6, IL-10, caspase 3, BCL-2 and GAPDH (dilution 1:1000; Beyotime Biotechnology) at 4°C overnight.

Techniques: Western Blot, Control