Journal: International Journal of Molecular Sciences

Article Title: MicroRNA-766-3p Contributes to Anti-Inflammatory Responses through the Indirect Inhibition of NF-κB Signaling

doi: 10.3390/ijms20040809

Figure Lengend Snippet: Generalizability of the suppression of inflammatory stimuli in miR-766-3p-transfected cells. ( A ) NHMCs were transfected with miRNA mimics (30 nM) and exposed to TNF-α or IL-1β for 24 h. The expression of the indicated genes was examined by a qPCR. ( B , C ) C28/I2 cells ( B ) and NHMCs ( C ) were co-transfected with pNF-κB-Luc along with miRNA mimics (C28/I2: 5 nM, NHMC: 30 nM). After incubation, cells were treated with TNF-α or IL-1β for 6 h and subjected to a luciferase assay to evaluate the activity of NF-κB. The luciferase activity was normalized by the number of viable cells and then normalized to the respective values in the vehicle samples (upper bar graphs). The lower scatterplots show the frequency of NF-κB inhibition by miR-766-3p. Assays were performed in quadruplicate. Data are expressed as the mean ± SEM. Asterisks indicate statistically significant differences ( p < 0.05).

Article Snippet: The human synovial fibroblast cell line MH7A was obtained from Riken Cell Bank (Ibaraki, Japan) [ ], the human chondrocyte cell line C28/I2 was obtained from Merck Millipore (Darmstadt, Germany), and normal human mesangial cells (NHMCs) were obtained from Lonza (Basel, Switzerland).

Techniques: Transfection, Expressing, Incubation, Luciferase, Activity Assay, Inhibition

Journal: Biochemical and biophysical research communications

Article Title: Effect of the PTHrP(1–34) analog abaloparatide on inducing chondrogenesis involves inhibition of intracellular reactive oxygen species production

doi: 10.1016/j.bbrc.2019.01.049

Figure Lengend Snippet: Mouse limb bud mesenchymal stem cells in micromass culture spontaneously undergo the entire process of differentiation. Representative of three independent experiments performed with similar results. A. Alcian blue staining on days 3 and 6. Bar: 2 mm. B. Col X expression by fluorescent staining on days 7 and 14 was shown in green and nuclear staining was shown in blue. Bar, 100 μm. Arrowheads indicate hypertrophic cells. C. Chondrocyte apoptosis occurs on day 21. Arrowheads show dark brown TUNEL-positive apoptotic cells. Normal nuclei are light green in color (methyl green counterstain). Bar: 20 μm. D. Cartilaginous tissue were stained with silver nitrate solution under UV light. Mineralized nodules were seen as dark brown to black spots. Bar, 100 μm.

Article Snippet: Mouse limb bud mesenchymal stem cells in micromass culture spontaneously undergo chondrogenesis, chondrocyte hypertrophy, apoptosis and terminal differentiation The cell model of mouse limb bud mesenchymal stem cells in micromass culture was used to examine different stages of mesenchymal cell differentiation.

Techniques: Staining, Expressing, TUNEL Assay

Journal: Biochemical and biophysical research communications

Article Title: Effect of the PTHrP(1–34) analog abaloparatide on inducing chondrogenesis involves inhibition of intracellular reactive oxygen species production

doi: 10.1016/j.bbrc.2019.01.049

Figure Lengend Snippet: ABL enhances mesenchymal stem cell chondrogenic differentiation. A. The confluence of mouse primary articular chondrocytes from newborn mice is shown in left panel. ABL and PTH(1–34) stimulated cAMP formation (middle panel) and intracellular calcium (right panel) in primary articular chondrocytes. B. Representative of three independent experiments performed with similar results for Alcian blue staining on day 5. Bar: 2 mm (upper panel). Quantitative assessment of cartilage nodule number (lower, left) and intensity of Alcian blue staining (lower right). Data are presented as mean ± SEM. n = 4. *, p < 0.05, **, p < 0.01, versus vehicle control.

Article Snippet: Mouse limb bud mesenchymal stem cells in micromass culture spontaneously undergo chondrogenesis, chondrocyte hypertrophy, apoptosis and terminal differentiation The cell model of mouse limb bud mesenchymal stem cells in micromass culture was used to examine different stages of mesenchymal cell differentiation.

Techniques: Staining, Control

Journal: Biochemical and biophysical research communications

Article Title: Effect of the PTHrP(1–34) analog abaloparatide on inducing chondrogenesis involves inhibition of intracellular reactive oxygen species production

doi: 10.1016/j.bbrc.2019.01.049

Figure Lengend Snippet: ABL inhibits ROS production in limb bud-derived mesenchymal cells. Mesenchymal stem cells (3 × 105 cells in 15 μl) were placed in the center of 24-well plate on day 0. On day 5, the cultures were serum-starved for 2 h and then loaded with DCF-DA (150 μM) for 10 min. After loading, ABL (10 nM), PTH(1 –34) (10 nM), NAC (1 mM) or vehicle was added to the cultures for 2 h. A. The fluorescence imaging was recorded using EVOS FL Auto Cell Imaging System and representative of three independent experiments performed with similar results is shown. Bar: 1 mm. B. DCF fluorescence intensity was measured, n = 5. *, p < 0.05, **, p < 0.01, versus vehicle control. C. Mesenchymal cells in micromass were treated with 10 nM ABL, 10 nM PTH(l-34), 1 mM NAC, or vehicle from day 0 to day 4. The mRNA expression of PTHtR was measured on days 1,3, and 5 by quantitative real time PCR. The data are presented as fold change and summarized as the means ± SEM. n = 4. a, p < 0.05, b, p < 0.01 versus vehicle on d1; c, p < 0.05, d, p < 0.01 versus vehicle on d3; e, p < 0.05, f. p < 0.01 versus vehicle on d5.

Article Snippet: Mouse limb bud mesenchymal stem cells in micromass culture spontaneously undergo chondrogenesis, chondrocyte hypertrophy, apoptosis and terminal differentiation The cell model of mouse limb bud mesenchymal stem cells in micromass culture was used to examine different stages of mesenchymal cell differentiation.

Techniques: Derivative Assay, Fluorescence, Imaging, Control, Expressing, Real-time Polymerase Chain Reaction