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: The Journal of Clinical Investigation

Article Title: Reprogramming dysfunctional CD8 + T cells to promote properties associated with natural HIV control

doi: 10.1172/JCI157549

Figure Lengend Snippet: Total CD8 + T cells from individuals without HIV were treated with medium, vehicle (Veh.) control, or the GSK3 inhibitor (inh), followed by incubation under basal conditions or with anti-CD3/anti-CD28 stimulation for 48 hours. ( A and B ) Analysis of CD8 + T cell subpopulations in unstimulated cells ( n = 4). ( C ) Fold change of CD8 + T cell subpopulations upon vehicle control or GSK3 inhibitor treatment, relative to the medium alone condition ( n = 4). ( D ) Expression of TCF-1 in CD8 + T cell subsets ( n = 4). ( E ) Fold change in the expression of the indicated markers induced by anti-CD3/anti-CD28 antibody stimulation relative to unstimulated cells ( n = 5). ( F ) Analysis of dead cells by Aqua LIVE/DEAD + staining (Aqua L/D) among total and T-bet + CD8 + T cells, and fold change in dead CD8 + T cells induced by anti-CD3/anti-CD28 stimulation relative to the unstimulated (Unstim.) condition ( n = 5). ( G ) Frequencies of granzyme B + (GZMB + ), IL-2 + , IFN-γ + , and TNF-α + CD8 + T cells after anti-CD3/anti-CD28 stimulation. ( H ) Expression of 1 to 4 functions in CD8 + T cells ( n = 5). * P < 0.05, by Dunn’s test ( B ) and Wilcoxon test ( C , D , and F – H ). Data obtained from 2 ( A – F ) or 3 ( G and H ) independent experiments are shown.

Article Snippet: Purified CD8 + T cells were stained with the LIVE/DEAD Fixable Aqua Dead Cell Stain kit and the following antibodies: anti–CD3 Alexa Fluor 700, anti–CD8 APC Cy7, anti–CCR7 PE Cy7, anti–CD45RA BV421, and anti–CD27 PE (all from BD Biosciences).

Techniques: Incubation, Expressing, Staining