Journal: PeerJ

Article Title: Hyperoside alleviates doxorubicin-induced myocardial cells apoptosis by inhibiting the apoptosis signal-regulating kinase 1/p38 pathway

doi: 10.7717/peerj.15315

Figure Lengend Snippet: (A) Representative fluorescence labeling images for terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) (green) in HL-1 cells. Scale bars, 50 µm. (B) Quantitative analysis of TUNEL-positive cells depicted by bar graph ( n = 3). (C) Representative immunofluorescence labeling images of cleaved caspase-3 (red) in HL-1 cells. Scale bars, 50 µm. (D) Quantitative analysis of cleaved caspase-3 depicted by bar graph ( n = 3). *** P < 0.001. Data are expressed as mean ± standard error of the mean (SEM) from three independent experiments.

Article Snippet: One-step terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) apoptosis assay kit, reactive oxygen species (ROS) assay kit and malondialdehyde (MDA) content assay kit purchased from Beyotime Biotech Inc (Shanghai, China).

Techniques: Fluorescence, Labeling, End Labeling, TUNEL Assay, Immunofluorescence

Journal: PeerJ

Article Title: Hyperoside alleviates doxorubicin-induced myocardial cells apoptosis by inhibiting the apoptosis signal-regulating kinase 1/p38 pathway

doi: 10.7717/peerj.15315

Figure Lengend Snippet: (A) CCK-8 assayed the effect of DOX on the survival rate of MDA-MB-231 cells. (B) CCK-8 assayed the effects of hyperoside on the survival rate of MDA-MB-231 cells induced by DOX. (C) Representative fluorescence labeling images for terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) (green) in MDA-MB-231 cells. Scale bars, 50 µm. (D) Quantitative analysis of TUNEL-positive cells depicted by bar graph ( n = 3). *** P < 0.001, ** P < 0.01, * P < 0.05. Data are expressed as mean ± standard error of the mean (SEM) from three independent experiments.

Article Snippet: One-step terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) apoptosis assay kit, reactive oxygen species (ROS) assay kit and malondialdehyde (MDA) content assay kit purchased from Beyotime Biotech Inc (Shanghai, China).

Techniques: CCK-8 Assay, Fluorescence, Labeling, End Labeling, TUNEL Assay

Journal: Cell Death & Disease

Article Title: STAT6 deficiency ameliorates Graves' disease severity by suppressing thyroid epithelial cell hyperplasia

doi: 10.1038/cddis.2016.398

Figure Lengend Snippet: STAT6 promoted TEC growth by inducing the expression of Bcl-xL and cyclin D1 EAGD was generated in WT and STAT6 −/− mice by injection with TSHR-289-expressing adenovirus as shown in . Mice were euthanized 4 weeks after three injections, and the thyroid glands were obtained. IF and IHC were used to assess the growth of TECs in the different groups. ( a ) Thyroid gland sections from WT and STAT6 −/− mice were assayed for TUNEL positivity (green) and stained with DAPI (blue) to determine the degree of TEC apoptosis. Apoptosis was measured in eight mice from each group. Representative staining is shown in the left panel. Arrows point to TUNEL-positive TECs. Magnification: × 200. IF was blindly scored by counting positive cells in 10 HPFs (right panel). ( b ) Ki67 was also measured in eight mice from each group by IHC staining. Representative Ki67 staining in each group is shown in the left panel. Arrow points to Ki67-positive TECs. Magnification: × 100; × 400 (left panel). IHC was blindly scored by counting positive cells in 10 HPFs (right panel). ( c ) Individual thyroid glands were obtained and homogenized. RNA and protein were then extracted from the homogenates. The mRNA expression of Bcl-xL ( c ) and cyclin D1 ( d ) was examined by quantitative real-time PCR analysis. ( e ) Bcl-xL and cyclin D1 protein levels were measured by western blotting. Immunoblot analysis of Bcl-xL and cyclin D1 levels in the individual thyroid gland were obtained from their relative band densities. GAPDH was used as the loading control. We analysed more than three individual mouse thyroid glands in every experiment. Independent experiments were repeated four times and a representative figure is shown. The data shown are the means±S.E.M. of one experiment with eight mice. Three replicates showed similar results. Scale bars in A: (× 200): 50 μm. Scale bars in B (× 100): 200 μm; scale bars (× 400): 50 μm. * P <0.05, ** P <0.01 and *** P <0.001

Article Snippet: The apoptotic TECs were stained with a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) assay (Apoptosis Detection Kit, Vazyme, China) according to the manufacturer's protocol and were analysed by the Olympus microscope camera system.

Techniques: Expressing, Generated, Injection, TUNEL Assay, Staining, Immunohistochemistry, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cell Death & Disease

Article Title: STAT6 deficiency ameliorates Graves' disease severity by suppressing thyroid epithelial cell hyperplasia

doi: 10.1038/cddis.2016.398

Figure Lengend Snippet: Phosphorylation of STAT6 is activated by IL-4 Nthy-ori 3-1 cells were stimulated by IL-4, and cells were harvested at different time points. ( a ) p-STAT6 was measured by western blotting (left panel). Immunoblot analysis of the ratio of pSTAT6/STAT6 in Nthy-ori 3-1 cells with their relative band densities (right panel). ( b ) Bcl-xL and cyclin D1 were measured by western blotting (left panel). Immunoblot analysis of Bcl-xL (middle panel) and cyclin D1 (right panel) with their relative band densities. GAPDH was used as a loading control. ( c ) Cells were stained by PI and annexin V to detect cell death and apoptosis by flow cytometry. The gating strategy is shown in the left two panels. The bar graphs show the percentage of annexin V + cells (right panel). ( d ) Cell viability was tested by a CCK8 kit. ( e ) p-STAT6 was measured by western blotting (left panel). Immunoblot analysis of the ratio of pSTAT6/STAT6 in Nthy-ori 3-1 cells with their relative band densities (right panel). ( f ) Bcl-xL and cyclin D1 were measured by western blotting (left panel). Immunoblot analysis of Bcl-xL (middle panel) and cyclin D1 (right panel) with their relative band densities. GAPDH was used as a loading control. ( g ) Cells were stained with PI and annexin V to determine cell death and apoptosis by flow cytometry. The bar graphs show the percentages of annexin V + cells ( h ) Cell viability was analysed by a CCK8 kit. The bar graph values are the means±S.E.M. of four independent experiments, each performed in duplicate. * P <0.05, ** P <0.01 and *** P <0.001

Article Snippet: The apoptotic TECs were stained with a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) assay (Apoptosis Detection Kit, Vazyme, China) according to the manufacturer's protocol and were analysed by the Olympus microscope camera system.

Techniques: Western Blot, Staining, Flow Cytometry