Journal: Oncology Reports

Article Title: Andrographolide suppresses aerobic glycolysis and induces apoptotic cell death by inhibiting pyruvate dehydrogenase kinase 1 expression

doi: 10.3892/or.2023.8509

Figure Lengend Snippet: AG induces more apoptotic cell death in H292 cells than A549 cells. (A and B) A549 and H292 cells were treated with various concentrations of AG (20, 50 and 75 µM) for 6 h, labeled with Annexin V-fluorescein isothiocyanate and propidium iodide, and analyzed via flow cytometry. (C) Histogram showing the percentage of apoptotic cells. Apoptotic cells were quantified by combining Q2 (late-stage of apoptotic cells, PI + /Annexin V + ) and Q3 (early-stage of apoptotic cells, PI − /Annexin V + ) regions after FACS analysis. AG significantly increased the apoptotic cell death in H292 cells, but not in A549 cells, in a dose-dependent manner. (D) To determine the protein levels of PARP, cleaved caspase-3, and cleaved caspase-9 via western blot analysis, A549 and H292 cells were treated with AG at the indicated dose for 24 h. Levels of all apoptotic marker proteins were significantly increased by AG treatment in H292 cells and only slightly increased in A549 cells. Data are represented as the mean ± SD compared with the control from three independent experiments. ***P<0.001 vs. control. AG, andrographolide; PARP, cleaved poly (ADP ribose) polymerase.

Article Snippet: Caspase-9 , Cell Signaling Technology, Inc. , 9508 , 1:1,000 in 2% skim milk.

Techniques: Labeling, Flow Cytometry, Western Blot, Marker

Journal: Cells

Article Title: Micrandilactone C, a Nortriterpenoid Isolated from Roots of Schisandra chinensis , Ameliorates Huntington’s Disease by Inhibiting Microglial STAT3 Pathways

doi: 10.3390/cells12050786

Figure Lengend Snippet: MC prevents neurodegeneration and apoptosis in the striatum after 3-NPA treatment. ( A – F ) At 24 h following the last (5th) 3-NPA treatment, striata from sham ( n = 5), 3-NPA ( n = 5), 3-NPA + MC (2.5 mg/kg/day; n = 5), and MC (2.5 mg/kg/day; n = 5) groups were subjected to histopathological and Western blot analyses to investigate the effect of MC (2.5 mg/kg/day, i.v.) on striatal cell death and apoptosis. ( A – C ) Representative photographs showing the level of striatal lesion by cresyl violet staining ( A ). MC significantly reduced the number of mice with striatal lesion ( B ) and the lesion area ( C ). Asterisk, S, and C indicate striatal lesions, normal striatum, and normal cortex, respectively. Scale bar = 100 μm. N.D., not detected. Data are expressed as mean ± SEM (Kruskal–Wallis; # p < 0.05 and ## p < 0.01 versus sham group; * p < 0.05 and ** p < 0.01 versus 3-NPA group). ( D , F , G ) Striata from all groups were analyzed by Western blot to determine the expression level of SDHA ( D ), cleaved caspase-9 ( G , H ), cleaved caspase-3 ( G , I ), and Bcl-2 ( G , J ). MC prevented the enhancement of these protein expressions. ( E , F ) Representative photographs ( E ) and quantified graph ( F ) showing the level of striatal cell death by FJC staining. MC significantly reduced the number of FJC (+) cells. Scale bar = 200 μm. Data are expressed as mean ± SEM (Kruskal–Wallis; ## p < 0.01 versus sham group; * p < 0.05 and ** p < 0.01 versus 3-NPA group).

Article Snippet: Briefly, 24 h after the last (5th) 3-NPA intoxication, the striatal proteins from all groups ( n = 5 per group) were incubated with primary antibodies, including mouse anti-succinate dehydrogenase complex subunit A (SDHA) (1:1000; Abcam, Cambridge, UK), rabbit an-ti-pro-caspase-3 (1:1000; Cell Signaling Technology), rabbit anti-cleaved caspase-3 (1:500; Cell Signaling Technology), rabbit anti-pro-caspase-9 (1:1000; Cell Signaling Technology), rabbit anti-cleaved caspase-9 (1:1000; Cell Signaling Technology), rabbit anti-B-cell lymphoma 2 (Bcl-2) (1:1000; Santa cruz Technology), rabbit anti-Iba-1 (1:500; WAKO), rabbit anti-phospho (p)-STAT3, STAT3 (1:500; Cell Signaling Technology), mouse anti-inducible nitric oxide synthases (iNOS) (1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-interleukin(IL)-1ß, IL-6, tumor necrosis factor-α (TNF-α) (1:1000; Cell Signaling Technology), mouse anti-neuronal nuclear protein (NeuN) (1:2000; Millipore), mouse anti-HTT ( clone mEM48; 1:500; Millipore), and mouse anti-HTT (clone 2Q75; 1:500; LifeSpan BioSciences, Seattle, WA, USA) antibodies.

Techniques: Western Blot, Staining, Expressing

Journal: Cells

Article Title: Micrandilactone C, a Nortriterpenoid Isolated from Roots of Schisandra chinensis , Ameliorates Huntington’s Disease by Inhibiting Microglial STAT3 Pathways

doi: 10.3390/cells12050786

Figure Lengend Snippet: MC prevents neurodegeneration and apoptosis in the striatum after 3-NPA treatment. ( A – F ) At 24 h following the last (5th) 3-NPA treatment, striata from sham ( n = 5), 3-NPA ( n = 5), 3-NPA + MC (2.5 mg/kg/day; n = 5), and MC (2.5 mg/kg/day; n = 5) groups were subjected to histopathological and Western blot analyses to investigate the effect of MC (2.5 mg/kg/day, i.v.) on striatal cell death and apoptosis. ( A – C ) Representative photographs showing the level of striatal lesion by cresyl violet staining ( A ). MC significantly reduced the number of mice with striatal lesion ( B ) and the lesion area ( C ). Asterisk, S, and C indicate striatal lesions, normal striatum, and normal cortex, respectively. Scale bar = 100 μm. N.D., not detected. Data are expressed as mean ± SEM (Kruskal–Wallis; # p < 0.05 and ## p < 0.01 versus sham group; * p < 0.05 and ** p < 0.01 versus 3-NPA group). ( D , F , G ) Striata from all groups were analyzed by Western blot to determine the expression level of SDHA ( D ), cleaved caspase-9 ( G , H ), cleaved caspase-3 ( G , I ), and Bcl-2 ( G , J ). MC prevented the enhancement of these protein expressions. ( E , F ) Representative photographs ( E ) and quantified graph ( F ) showing the level of striatal cell death by FJC staining. MC significantly reduced the number of FJC (+) cells. Scale bar = 200 μm. Data are expressed as mean ± SEM (Kruskal–Wallis; ## p < 0.01 versus sham group; * p < 0.05 and ** p < 0.01 versus 3-NPA group).

Article Snippet: Briefly, 24 h after the last (5th) 3-NPA intoxication, the striatal proteins from all groups ( n = 5 per group) were incubated with primary antibodies, including mouse anti-succinate dehydrogenase complex subunit A (SDHA) (1:1000; Abcam, Cambridge, UK), rabbit an-ti-pro-caspase-3 (1:1000; Cell Signaling Technology), rabbit anti-cleaved caspase-3 (1:500; Cell Signaling Technology), rabbit anti-pro-caspase-9 (1:1000; Cell Signaling Technology), rabbit anti-cleaved caspase-9 (1:1000; Cell Signaling Technology), rabbit anti-B-cell lymphoma 2 (Bcl-2) (1:1000; Santa cruz Technology), rabbit anti-Iba-1 (1:500; WAKO), rabbit anti-phospho (p)-STAT3, STAT3 (1:500; Cell Signaling Technology), mouse anti-inducible nitric oxide synthases (iNOS) (1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-interleukin(IL)-1ß, IL-6, tumor necrosis factor-α (TNF-α) (1:1000; Cell Signaling Technology), mouse anti-neuronal nuclear protein (NeuN) (1:2000; Millipore), mouse anti-HTT ( clone mEM48; 1:500; Millipore), and mouse anti-HTT (clone 2Q75; 1:500; LifeSpan BioSciences, Seattle, WA, USA) antibodies.

Techniques: Western Blot, Staining, Expressing