Journal: PLoS ONE

Article Title: Stromal Cell-Derived Factor 1 Regulates the Actin Organization of Chondrocytes and Chondrocyte Hypertrophy

doi: 10.1371/journal.pone.0037163

Figure Lengend Snippet: Cytoskeleton of hypertrophic chondrocytes in the humeri. A : Frozen sections of the humeri isolated from SDF-1 −/− (KO), and wild-type (WT) mice at E15.5, were fixed with paraformaldehyde and stained with Rhodamine-Phalloidin for actin filaments and SYBR Green for nuclei. Left panel, Rhodamine-Phalloidin; middle panel, merge of Rhodamine-Phalloidin and SYBR Green; right panel, differentiation interference image. Representative fields are shown. B : The intensity of actin filament staining (F-actin) was quantified with Image J software. Values were normalized to F-actin of WT chondrocytes. C : The ratios of the short axis to the long axis (1/Ellipticity) in chondrocytes were calculated with Image J. Values are the mean and s.e.m. of more than three separate experiments; *, P

Article Snippet: In addition, the influence of genetic background of SDF-1−/− mice cannot completely be removed.

Techniques: Isolation, Mouse Assay, Staining, SYBR Green Assay, Software

Journal: PLoS ONE

Article Title: Stromal Cell-Derived Factor 1 Regulates the Actin Organization of Chondrocytes and Chondrocyte Hypertrophy

doi: 10.1371/journal.pone.0037163

Figure Lengend Snippet: SDF-1 protein was expressed at the prehypertrophic and hypertrophic zones. The growth plate of 4-week-old mouse tibia (A) and endochondral callus of rib fracture (B) were stained with hematoxylin and eosin (HE), or immunohistochemically stained with antiSDF-1 antibody or IgG (control). Boxed areas in the panel are shown in a higher magnification (×200) in the right panels. The result is representative of three separate experiments; Scale bar, 200 µm.

Article Snippet: In addition, the influence of genetic background of SDF-1−/− mice cannot completely be removed.

Techniques: Staining

Journal: PLoS ONE

Article Title: Stromal Cell-Derived Factor 1 Regulates the Actin Organization of Chondrocytes and Chondrocyte Hypertrophy

doi: 10.1371/journal.pone.0037163

Figure Lengend Snippet: Lack of SDF-1 delays the growth metatarsals in organ culture. A : The metatarsal bones were harvested from SDF-1 −/− (KO), SDF-1+/− (hetero), and wild-type (WT) mice at E15.5, cultured for 7 days, and the total length was measured at day 1, 3, 5, and 7. The length of metatarsal bones was calculated by day 1 as controls. B : Calcified spots (arrows) were counted in KO and WT metatarsals at day 1 and 7. C : Metatarsal bones were stained with hematoxylin and eosin (upper panel), or von Kossa (lower panel); Scale bar, 200 µm. D : The expression levels of type X collagen (Col X) and Sox9 of cultured metatarsal bones from E15.5 WT and KO mice at day 3 and 10 were quantified by real-time PCR. E : Primary chondrocytes isolated from WT and KO mice were cultured for 3 weeks in chondrogenic medium supplemented with insulin. The expression levels of Col X and Sox9 were quantified by real-time PCR. Values are the mean and s.e.m. of more than three independent experiments; *, P

Article Snippet: In addition, the influence of genetic background of SDF-1−/− mice cannot completely be removed.

Techniques: Organ Culture, Mouse Assay, Cell Culture, Staining, Expressing, Real-time Polymerase Chain Reaction, Isolation

Journal: PLoS ONE

Article Title: Stromal Cell-Derived Factor 1 Regulates the Actin Organization of Chondrocytes and Chondrocyte Hypertrophy

doi: 10.1371/journal.pone.0037163

Figure Lengend Snippet: SDF-1 simulates actin assembly and stress fiber formation in a time-dependent manner. Primary chondrocytes from wild-type mice were treated with SDF-1 (100 ng/ml) as indicated. Cells were then fixed with paraformaldehyde and stained with Rhodamine-Phalloidin for actin filaments and 4′,6-diamidino-2-phenylindole for nuclei; Scale bar, 20 µm.

Article Snippet: In addition, the influence of genetic background of SDF-1−/− mice cannot completely be removed.

Techniques: Mouse Assay, Staining

Journal: PLoS ONE

Article Title: Stromal Cell-Derived Factor 1 Regulates the Actin Organization of Chondrocytes and Chondrocyte Hypertrophy

doi: 10.1371/journal.pone.0037163

Figure Lengend Snippet: SDF-1/CXCR4 pathway controls actin cytoskeleton. A : Treatment with SDF-1 increased the actin filament density in primary chondrocytes. Primary chondrocytes from wild-type mice were treated with SDF-1 (100 ng/ml), or SDF-1 and CXCR4 specific antagonist, TF14016 (100 µM) or pertussis toxin (PTX, 100 ng/ml) for 60 m and stained with Rhodamine-Phalloidin. B : The F-actin content in chondrocytes was analyzed with Image J software. All values were normalized to F-actin of chondrocytes treated with conditioned medium only. C : Actin cytoskeleton of primary chondrocyte from SDF-1 −/− (KO), and wild-type (WT) mice. D : The F-actin content in chondrocytes was quantified with Image J software. All values were normalized to the F-actin of wild-type chondrocyte. Values are the mean and s.e.m. of more than three separate experiments; *, P

Article Snippet: In addition, the influence of genetic background of SDF-1−/− mice cannot completely be removed.

Techniques: Mouse Assay, Staining, Software

Journal: PLoS ONE

Article Title: Stromal Cell-Derived Factor 1 Regulates the Actin Organization of Chondrocytes and Chondrocyte Hypertrophy

doi: 10.1371/journal.pone.0037163

Figure Lengend Snippet: Growth of the embryonic humeri of SDF-1 −/− mice delays. A : The embryonic humeri of wild-type (WT) and SDF-1 −/− (KO) mice. Specimens were processed to paraffin-embedded sections and stained with hematoxylin, eosin (lower panels) and alcian blue (upper panels). B : Length of the humeri. The total length of the humeri and the ratios of the proliferating, the hypertrophic, or the calcified zone in each day were calculated as described in Methods section. Values are the mean and standard deviation of more than three independent experiments; *, P

Article Snippet: In addition, the influence of genetic background of SDF-1−/− mice cannot completely be removed.

Techniques: Mouse Assay, Staining, Standard Deviation

Journal: International Journal of Molecular Medicine

Article Title: Neuroprotective effects of Scrophularia buergeriana extract against glutamate-induced toxicity in SH-SY5Y cells

doi: 10.3892/ijmm.2019.4139

Figure Lengend Snippet: Effects of Scrophularia buergeriana extract (SBE) on apoptotic protein expression levels in SH-SY5Y cells. (A) The expression of phosphorylated (p)-p38, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3 and cleaved poly (adenosine diphosphate (ADP)-ribose) polymerase (PARP) was measured by western blot analysis. (B) The density of the protein bands was quantified and calculated using ImageJ software. p-p38 protein expression levels were normalized to those of p38, and other protein expression levels were normalized to those of β-actin. The data are expressed as the means ± SEM of independent experiments (n=3). * P

Article Snippet: After washing, the membranes were incubated at 4°C overnight with the following appropriate antibodies: SOD1 (1:1,000; cat. no. sc-515404), SOD2 (1:1,000; cat. no. sc-137254) (both from Santa Cruz Biotechnology, Santa Cruz, CA, USA), GPx-1 (1:1,000; cat. no. 3206), p-p38 (1:1,000; cat. no. 9211), p38 (1:1,000; cat. no. 8690), Bax (1:1,000; cat. no. 2772), Bcl-2 (1:1,000; cat. no. 3498), cleaved caspase-3 (1:1,000; cat. no. 9664), cleaved PARP (1:1,000; cat. no. 5625) and β-actin (1:2,000; cat. no. 8457) (all from Cell Signaling Technology, Danvers, MA, USA) washed 3 times with TBST, and incubated with horseradish peroxidase-conjugated secondary antibodies: goat anti-rabbit (1:1,000; cat. no. SA002), goat anti-mouse (1:1,000; cat. no. SA001) (both from genDEPOT, Katy, TX, USA) for 1 h at room temperature.

Techniques: Expressing, Western Blot, Software

Journal: International Journal of Molecular Medicine

Article Title: Neuroprotective effects of Scrophularia buergeriana extract against glutamate-induced toxicity in SH-SY5Y cells

doi: 10.3892/ijmm.2019.4139

Figure Lengend Snippet: Schematic view of Scrophularia buergeriana extract (SBE) regulating reactive oxygen species (ROS) levels and the apoptotic signaling cascade. Excessive glutamate caused ROS generation and induces neuronal cell apoptosis. The generated ROS triggers p38 and Bcl-2-associated X protein (Bax) protein and damage to mitochondrial function. The activated caspase-3 and poly (adenosine diphosphate (ADP)-ribose) polymerase (PARP) proteins result in apoptosis. SBE treatment reduces the production of ROS by regulating the expression of enzymes, including superoxide dismutase (SOD)1, SOD2 and glutathione peroxidase-1 (GPx-1) and also inhibits the apoptotic pathway.

Article Snippet: After washing, the membranes were incubated at 4°C overnight with the following appropriate antibodies: SOD1 (1:1,000; cat. no. sc-515404), SOD2 (1:1,000; cat. no. sc-137254) (both from Santa Cruz Biotechnology, Santa Cruz, CA, USA), GPx-1 (1:1,000; cat. no. 3206), p-p38 (1:1,000; cat. no. 9211), p38 (1:1,000; cat. no. 8690), Bax (1:1,000; cat. no. 2772), Bcl-2 (1:1,000; cat. no. 3498), cleaved caspase-3 (1:1,000; cat. no. 9664), cleaved PARP (1:1,000; cat. no. 5625) and β-actin (1:2,000; cat. no. 8457) (all from Cell Signaling Technology, Danvers, MA, USA) washed 3 times with TBST, and incubated with horseradish peroxidase-conjugated secondary antibodies: goat anti-rabbit (1:1,000; cat. no. SA002), goat anti-mouse (1:1,000; cat. no. SA001) (both from genDEPOT, Katy, TX, USA) for 1 h at room temperature.

Techniques: Generated, Expressing