Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Paeoniflorin Attenuates Dexamethasone-Induced Apoptosis of Osteoblast Cells and Promotes Bone Formation via Regulating AKT/mTOR/Autophagy Signaling Pathway

doi: 10.1155/2021/6623464

Figure Lengend Snippet: Effect of paeoniflorin on DEX-induced apoptosis in MC3T3-E1 cells. (a, b) The extent of apoptosis of MC3T3-E1 cells was detected by flow cytometry after Annexin V-FITC/PI double staining. The apoptotic rate was measured by flow cytometric analysis. Data are presented as the mean ± SD. ∗∗ P < 0.01 vs. control; # P < 0.05 vs. DEX. (c) MC3T3-E1 cells were stained by Hoechst to show the extent of apoptosis after interventions of DEX and paeoniflorin. (d) The expressions of osteogenic protein-Runx2 (runt-related transcription factor2) and apoptotic proteins such as Bcl-2 (B cell leukemia 2) and Bax (BCL2-associated X protein) were measured by western blot. ∗ P < 0.05, ∗∗ P < 0.01 vs. control; # P < 0.05 vs. DEX; ## P < 0.01 vs. DEX. (e) Protein levels of caspase-3 and cleaved caspase-3 were determined by western blot analysis, shown in quantitative analysis. Data are presented as the mean ± SD. ∗∗ P < 0.01 vs. control; # P < 0.05 vs. DEX.

Article Snippet: For Hoechst staining, 10 μ L Hoechst live cell staining solution (Beyotime Institute of Biotechnology) was added to the culture medium and mixed gently.

Techniques: Flow Cytometry, Double Staining, Control, Staining, Western Blot

Journal: Experimental and Therapeutic Medicine

Article Title: Scutellaria barbata D. Don inhibits growth and induces apoptosis by suppressing IL-6-inducible STAT3 pathway activation in human colorectal cancer cells

doi: 10.3892/etm.2015.2692

Figure Lengend Snippet: Effect of EESB on the HT-29 cell cycle. Cells were pretreated with various concentration of EESB for 1 h, followed by stimulation with 10 ng/ml IL-6 for 24 h. (A) Cells were stained with propidium iodide and analyzed by fluorescence-activated cell sorting. The proportion of DNA in the S phase was calculated using ModFit LT version 3.0 software. (B) Quantification of fluorescence-activated cell sorting analysis. The data shown are averages with standard deviation from 3 independent experiments. #P<0.05 vs. cells treated with IL-6 but not EESB. EESB, ethanol extract of Scutellaria barbata D. Don; IL-6, interleukin-6.

Article Snippet: The HT-29 cell cycle progression was determined through flow cytometric analysis using a propidium iodide (PI) staining cell cycle assay kit (BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Concentration Assay, Staining, Fluorescence, FACS, Software, Standard Deviation

Journal: Experimental and Therapeutic Medicine

Article Title: Scutellaria barbata D. Don inhibits growth and induces apoptosis by suppressing IL-6-inducible STAT3 pathway activation in human colorectal cancer cells

doi: 10.3892/etm.2015.2692

Figure Lengend Snippet: Effect of EESB on HT-29 cell apoptosis. Cells were pretreated with various concentration of EESB for 1 h, followed by stimulation with 10 ng/ml IL-6 for 24 h. (A) Cells were collected and stained with Annexin V/PI, followed by fluorescence-activated cell sorting analysis. Double-negative stained cells indicate the live cell population; Annexin V-positive/PI-negative stained cells and Annexin V/PI double-positive stained cells represent early and late apoptosis, respectively; Annexin V-negative and PI-positive stained cells show dead cells. (B) Quantification of fluorescence-activated cell sorting analysis. The data shown are averages with standard deviation from 3 independent experiments. #P<0.05 vs. cells treated with IL-6 but not EESB. EESB, ethanol extract of Scutellaria barbata D. Don; IL-6, interleukin-6; UL, upper left; UR, upper right; LR, lower right; LL, lower left; PI, propidium iodide; FITC, fluorescein isothiocyanate.

Article Snippet: The HT-29 cell cycle progression was determined through flow cytometric analysis using a propidium iodide (PI) staining cell cycle assay kit (BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Concentration Assay, Staining, Fluorescence, FACS, Standard Deviation

Journal: Oncotarget

Article Title: A novel human truncated IL12rβ1-Fc fusion protein ameliorates experimental autoimmune encephalomyelitis via specific binding of p40 to inhibit Th1 and Th17 cell differentiation

doi:

Figure Lengend Snippet: Splenocytes, lymph nodes cells and CNS MNCs of tIL12rβ1/Fc- or vehicle-treated EAE mice at day 18 post-immunization (therapeutic protocol) were isolated. A. Supernatants derived from splenocytes, lymph nodes cells and CNS MNCs reactivated with MOG 35–55 (20 μg/mL) for 24 h were analyzed for the level of p40 (mean ± SD; n = 5). B. The percentage of Th1, Th1/Th17 and Th17 cells in the CD4 + lymphocyte gate were analyzed by intracellular staining of IFN-γ and IL-17, following stimulation with MOG 35–55 (20 μg/mL) for 24 h. C. The Th17:Th1 ratios in CNS, LNs and Spleens were observed for each group. Correlation of Th17:Th1 ratios with vehicle treatment and that with tIL12rβ1/Fc treatment was significant ( p < 0.05) ( n = 4). D. Percentages of cells positive expression with these antigens in spleen, LN or CNS are expressed as mean ± SD ( n = 4). E . The frequency of regulatory T cells in the CD4 + lymphocyte gate were analyzed by intracellular staining of Foxp3 following stimulation with MOG 35–55 (20 μg/mL) for 24 h. Percentage of Foxp3 + cells in spleen is expressed as mean ± SD ( n = 4). * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The detection of regulatory T (Treg) cells was carried out using the Regulatory T Cell Staining kit (BD, USA), according to the manufacturer's instructions.

Techniques: Isolation, Derivative Assay, Staining, Expressing