Journal: Life science alliance

Article Title: Mitochondrial impairment and intracellular reactive oxygen species alter primary cilia morphology.

doi: 10.26508/lsa.202201505

Figure Lengend Snippet: Figure 2. Reactive oxygen species, but not cells cycle and apoptosis, are in part responsible for decreased ciliation in IMCD3. (A) Quantification of Mitosox intensity in IMCD3 cells treated for 48 h with DMSO vehicle, 25 nM rotenone, or 50 nM oligomycin. Data are represented by mean + SD; n = 6. (B) Quantification of DCFDA intensity in IMCD3 cells treated as above. Data are represented by mean + SD; n = 5. (C) Quantification of cilia number in IMCD3 cells treated for 48 h with 25 nM of rotenone in the presence of antioxidants (Tiron + Vit.C 1 mM, retinoic acid 1 μM). Data are represented by mean + CI; n = 4. (D) Cell cycle analysis in IMCD3 cells treated as above. In the inset is displayed a representative image of cell cycle plot in control cells. Data are represented by mean + CI; n = 4. (E) Analysis of apoptotic and dead IMCD3 cells treated as above normalized per 100 events of DMSO treated cells. In the inset is displayed a representative plot of control cells. Staurosporin 6 μM has been added 2 h prior the trypsinization as positive control. Data are represented by mean + SEM; n ≥2. Legend: Black empty = DMSO control; black = rotenone 25 nM; red = oligomycin 50 nM; blue = staurosporine 6 μM; orange = Tiron + Vit.C 1 mM; pink = Retinoic acid 1 μM. Statistical significance between control versus treatments: P < 0.05 (*), <0.01 (**), or <0.001 (***). Rotenone versus oligomycin: P < 0.05 (#), <0.01 (##), or <0.001 (###).

Article Snippet: To measure apoptosis in living cells we used Annexin V-FITC Apoptosis Detection Kit (Abcam), according to the manufacturer’s instruction.

Techniques: Cell Cycle Assay, Control, Positive Control

Journal: ACS Omega

Article Title: Study on Antitumor Secondary Metabolites from Euphorbia ebracteolata Hayata: Structural Characterization, Network Pharmacology, Molecular Docking and In Vitro

doi: 10.1021/acsomega.5c04875

Figure Lengend Snippet: Effect of compound (±)- 1 on apoptosis and cell-cycle profiling in SMMC-7721 cells. Cells were exposed to varying concentrations of compound (±)- 1 (0, 0.1, 0.3, 0.6 μM) for 24 h. (A) Apoptosis of SMMC-7721 cells was assayed by flow cytometry. (B) Cell-cycle profiling was performed using a FACSCanto flow cytometer. Data and profiles for the G0/G1 and G2/M phases, analyzed with Diva software and ModFit LT ver. 3.0, are shown in the gray area. Cells in the S phase are represented as the shaded area. (C) The proportion of apoptotic cells was quantified and expressed as the mean ± SD of three independent experiments (*** p < 0.001). (D) Representative FACS histograms from three independent experiments.

Article Snippet: The Cell Cycle and Apoptosis Analysis Kit, Annexin V-FITC Apoptosis Detection Kit, Mitochondrial Membrane Potential and Apoptosis Detection Kit with Mito-Tracker Red CMXRos and Annexin V-FITC were obtained from Beyotime (Shanghai, China).

Techniques: Flow Cytometry, Software