Journal: Nucleic Acids Research

Article Title: Gli2 and MEF2C activate each other's expression and function synergistically during cardiomyogenesis in vitro

doi: 10.1093/nar/gkr1232

Figure Lengend Snippet: Gli2 and MEF2C are expressed during mES differentiation. ( A ) Schematic representation of cardiomyogenesis in D3 mES cells, as described in ‘Materials and Methods’ section. ( B–E ) mES cells were differentiated and examined (B) for MHC expression using MF20 antibodies on Day 9 of differentiation. Nuclei were stained with Hoechst, scale bar is 30 μM; (C and D) by QPCR analysis for the indicated genes on Days 0, 3, 6 and 9 of differentiation, n = 3. Error bars represent ± SEM. Primers are listed in . (C) Values were compared with Day 0 values and statistical significance was determined using ANOVA (* P < 0.05 and ** P < 0.01). (D) To determine statistical significance between all days analysed, ANOVA followed by post hoc Tukey HSD analysis was performed, * P < 0.05 and ** P < 0.01; n.s. = not significant.; (E) immunoblot analysis of total protein extracts using MEF2C- and Gli2-specific antibodies at the time shown. Arrows designate Gli2 or MEF2C protein band(s), and asterisk denotes non-specific binding of the Gli2 antibodies. α-Tubulin served as a loading control. Lanes were spliced out from the same autoradiogram as designated by vertical lines. Gli2 band densities were measured from one representative experiment using ImageJ program , normalized to α-tubulin and expressed as percent maximum; ( F ) immunoblot analysis of eye total protein extracts from E16 Gli2 +/− and Gli2 −/− mice using Gli2-specific antibodies. Asterisk denotes non-specific binding of the antibodies. β-Actin served as a loading control.

Article Snippet: D3 mES cells (ATCC, # CRL-1934) were maintained as described in ref. ( , ).

Techniques: Expressing, Staining, Western Blot, Binding Assay, Control

Journal: Nucleic Acids Research

Article Title: Gli2 and MEF2C activate each other's expression and function synergistically during cardiomyogenesis in vitro

doi: 10.1093/nar/gkr1232

Figure Lengend Snippet: Summary of gene expression changes in cell lines treated with or without DMSO

Article Snippet: D3 mES cells (ATCC, # CRL-1934) were maintained as described in ref. ( , ).

Techniques: Gene Expression

Journal: International Journal of Medical Sciences

Article Title: Controlling Osteogenesis and Adipogenesis of Mesenchymal Stromal Cells by Regulating A Circadian Clock Protein with Laser Irradiation

doi:

Figure Lengend Snippet: Intracellular location of (A) mCRY1 and (B) mPER2 proteins in MSCs 24 hr after laser irradiation. Cells were double-labeled with DAPI (blue, upper panel) and mCRY1 or mPER2 (red, center panel). The lower panel provides a merged image. mCRY1 and mPER2 localized to the cytoplasm prior to laser irradiation. After laser irradiation, proteins translocated to the nucleus. (C) mRNA levels of mCry1 in MSCs 24 hr after laser irradiation (100 mW/cm ) and in non-irradiated cells. Samples were normalized to mRps18 . The mRNA levels of mCry1 decreased after blue laser irradiation relative to non-irradiated cells. Scale bars = 30µm *, p<0.01: significant difference between the relative mRNA levels of laser irradiated MSCs and controls.

Article Snippet: The mouse MSCs cell line (KUSA-A) was purchased from RIKEN Bioresource Center, Japan, and cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal calf serum (FCS), 100 units/mL penicillin, and 0.1 mg/mL streptomycin at 37°C in a 5% CO 2 atmosphere.

Techniques: Irradiation, Labeling

Journal: International Journal of Medical Sciences

Article Title: Controlling Osteogenesis and Adipogenesis of Mesenchymal Stromal Cells by Regulating A Circadian Clock Protein with Laser Irradiation

doi:

Figure Lengend Snippet: (A) The beam profile of the blue laser (wave length; 405 nm, continuous wave). MSCs were irradiated for 180 sec at various intensities. (B) Histochemical analysis of laser irradiated MSCs. Alizarin red-S staining of irradiated MSCs (magnification: x50). At 5 days post-irradiation, calcium deposition had increased around the cells in a dose-dependent manner. Calcium phosphate deposition was evaluated by von Kossa staining (magnification: x50). At 5 days after treatment, staining increased with increased laser energy. The area expressing alkaline phosphatase (ALP) activity was stained (magnification: x50). Laser irradiated samples displayed immunopositive staining for osteocalcin, a marker of osteoblast differentiation (magnification: x100). Scale bars = 200 (for Alizarin red-S, von Kossa, and ALA staining) and 100µm (for osteocalcin immunostaining). (C) The quantitative calcium content increased after blue laser irradiation relative to non-irradiated cells. Calcium content increases varied with laser energy. *, p<0.01: significant difference between the calcium content of laser-irradiated MSCs and controls.

Article Snippet: The mouse MSCs cell line (KUSA-A) was purchased from RIKEN Bioresource Center, Japan, and cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal calf serum (FCS), 100 units/mL penicillin, and 0.1 mg/mL streptomycin at 37°C in a 5% CO 2 atmosphere.

Techniques: Irradiation, Staining, Expressing, Activity Assay, Marker, Immunostaining

Journal: International Journal of Medical Sciences

Article Title: Controlling Osteogenesis and Adipogenesis of Mesenchymal Stromal Cells by Regulating A Circadian Clock Protein with Laser Irradiation

doi:

Figure Lengend Snippet: (A) Staining with oil red O demonstrates that blue laser irradiation decreased adipogenesis relative to non-irradiated areas (magnification: x50). Higher magnification (x400) is shown in frame. Scale bars = 200 µm. As the nuclear localization of CRY proteins attenuates CLOCK- and BMAL1-driven transcription, laser irradiation may act 'molecular switch' for regulatory proteins by suppressing CRY transcription to limit the accumulation of lipid droplets in cells. (B) mRNA levels of PPARγ in MSCs 24 hr after laser irradiation (100 mW/cm 2 ) and in non-irradiated cells. Samples were normalized to mRps18 . The mRNA levels of PPARγ decreased after blue laser irradiation relative to non-irradiated cells. *, p<0.01: significant difference between the relative mRNA levels of laser irradiated MSCs and controls.

Article Snippet: The mouse MSCs cell line (KUSA-A) was purchased from RIKEN Bioresource Center, Japan, and cultured in Dulbecco's Modified Eagle's Medium (DMEM) containing 10% fetal calf serum (FCS), 100 units/mL penicillin, and 0.1 mg/mL streptomycin at 37°C in a 5% CO 2 atmosphere.

Techniques: Staining, Irradiation