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Image Search Results
Journal: Biomolecules
Article Title: Microglia Activated by Excess Cortisol Induce HMGB1 Acetylation and Neuroinflammation in the Hippocampal DG Region of Mice Following Cold Exposure
doi: 10.3390/biom9090426
Figure Lengend Snippet: Expression of mitogen-activated protein kinase (MAPK) signaling pathway components in BV2 cells after cortisol (CORT) treatment (200 μM) for 3 h, as assessed by western blotting. ( A ) Extracellular signal-regulated kinase (ERK), phospho- (p-)ERK (Thr202/Tyr204), c-Jun N-terminal kinase (JNK), p-JNK (Thr183/Tyr185), p38, p-p38 (Thr180/Tyr182), and β-actin expression. ( B ) Quantification of expression levels based on p-ERK/ERK, p-JNK/JNK, and p-p38/p38 ratios. Differences between control and CORT groups were evaluated by one-way analysis of variance. Results are expressed as the mean ± SD ( n = 3) of three independent experiments. *** p < 0.001.
Article Snippet: Approximately 30 μg of total protein was separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (0.22 and 0.45 μm; Millipore, Darmstadt, Germany) that were blocked for 1 h at room temperature in 5% nonfat milk in Tris-buffered saline containing 0.1% Tween 20 (TBST), which were then incubated overnight at 4 °C with antibodies against the following proteins: HMGB1 (#21865-1-AP, 1:3000), GFAP (#16825-1-AP, 1:2000), ionized calcium-binding adapter molecule (IBA)-1 (#10904-1-AP, 1:500), p65 (#10745-1-AP, 1:1000), silent mating type information regulation 2 homolog (SIRT)1 (#66292-1-Ig, 1:3000), brain-derived neurotrophic factor (BDNF); #16806-1-AP, 1:1000), IL-1β (#18985-1-AP, 1:1000), inducible nitric oxide synthase (iNOS) (#12987-1-AP, 1:1000), lamin B1 (#17168-1-AP, 1:3000), histone H3 (#60008-1-lg, 1:15,000), and β-actin (#14395-1-AP, 1:15,000) (all from
Techniques: Expressing, Western Blot