Journal: International Journal of Ophthalmology

Article Title: Netrin-1 promotes epithelium repair in corneal injury

doi: 10.18240/ijo.2020.02.02

Figure Lengend Snippet: A-B: Measured the cells apoptosis with flow cytometry in conjunction with annexin V-FITC/PI double staining. The values represent the mean percentages of apoptotic cells (aP<0.05). C. Quantization of the numbers of apoptotic cells. Every value represents the mean±SD, n=3. aP<0.05.

Article Snippet: An apoptosis assay kit with Hoechst 33342 and PI staining (Beyotime Institute of Biotechnology, Shanghai, China) was used to analyze apoptosis, in accordance with the instructions.

Techniques: Flow Cytometry, Double Staining

Journal: Cancer Research

Article Title: Increased Expression of Annexin A3 Is a Mechanism of Platinum Resistance in Ovarian Cancer

doi: 10.1158/0008-5472.can-09-3215

Figure Lengend Snippet: Figure 1. Annexin A3 expression is increased in platinum-resistant ovarian cancer cell lines and tumors from patients. A, the relative viability of the sensitive and resistant cell lines in the presence of cisplatin or carboplatin. Compared with their parent human ovarian cancer cell lines SKOV3 and A2780, platinum-selected SKOV3/Cis, A2780/Cis, SKOV3/Car, and A2780/Car are relatively resistant to cisplatin and carboplatin. B, intracellular staining of annexin A3 in SKOV3, A2780, and derived resistant cell lines. Anti–annexin A3 antibody and fluorescein-conjugated second antibody were used to visualize annexin A3 (green), and propidium iodide (PI) was used to stain cell nuclei (red). Annexin A3 is largely localized in cytoplasm, particularly the perinuclear area. The level of annexin A3 is increased in all the resistant cell lines. Bar, 20 μm. C, immunohistochemical staining of annexin A3 in various ovarian cancers. Top left, negative staining of endometrioid carcinoma; top right, positive staining of serous papillary cystadenocarcinoma. Bottom left, positive staining of transitional cell carcinoma; bottom right, positive staining of clear cell carcinoma. Bar, 100 μm. D, scores of annexin A3 immunohistochemical staining in ovarian cancer sections. Tumor sections were obtained from 21 chemosensitive and 21 chemoresistant patients. Analysis with Mann-Whitney U test shows that there is a significant difference between the two groups.

Article Snippet: Following incubation with goat anti-rabbit IgG antibodies conjugated with fluorescein (Santa Cruz Biotechnology), cell nuclei were stained with propidium iodide for 20 min.

Techniques: Expressing, Staining, Derivative Assay, Immunohistochemical staining, Negative Staining, MANN-WHITNEY

Journal: Journal of Inflammation Research

Article Title: Lycopus lucidus Turcz Exerts Neuroprotective Effects Against H 2 O 2 -Induced Neuroinflammation by Inhibiting NLRP3 Inflammasome Activation in Cortical Neurons

doi: 10.2147/JIR.S305031

Figure Lengend Snippet: LLT protects cortical neurons against H 2 O 2 -induced neuronal damage. ( A ) Schematic depiction of experimental design for primary cortical neurons ( B ) Cell viability assay results of cortical neurons treated with various concentrations of LLT without H 2 O 2 induction at 24 h, n = 3. ( C ) CCK-8 results of H 2 O 2 -treated cortical neurons treated with various concentrations of LLT at 24 h, n = 3. ( D ) The total number of live and dead cells calculated from cellular fluorescence images of each group. n = 10. ( E ) The percentage of apoptotic cells analyzed by flow cytometry of Annexin-V-FITC/PI-PE double staining. n = 5. ( F ) Fluorescence images of live and dead (red) cells for analysis of the neuroprotective effect of LLT at different concentrations on H 2 O 2 -treated cortical neurons. Scale bar = 200 µm. ( G ) Representative flow cytometric plot using Annexin-V/PI to confirm the mode of cell death. Blank = non-treatment, H 2 O 2 = H 2 O 2 only, LLT = H 2 O 2 +LLT extract. Significant differences are indicated as follows and were analyzed via one-way ANOVA with Tukey’s post-hoc test: # p < 0.001 vs the blank group; * p < 0.05, ** p < 0.01, and *** p < 0.001 vs the H 2 O 2 group.

Article Snippet: Apoptotic cell death was detected using an Annexin V-Fluorescein isothiocyanate (FITC) propidium iodide (PI)-phycoerythrin apoptosis detection kit (Abcam) as described previously.

Techniques: Viability Assay, CCK-8 Assay, Fluorescence, Flow Cytometry, Double Staining

Journal: Theranostics

Article Title: Role of damaged mitochondrial transfer in alpha-particle generator 212 Pb radiation-induced bystander effect

doi: 10.7150/thno.101922

Figure Lengend Snippet: Radiation-induced bystander effect triggered by alpha-particle 212 Bi. A) Decay scheme of alpha-particle generator 212 Pb. B) Schematic representation of the experimental design for the subsequent cytotoxic effect study. C, D, and E) The cytotoxic effect of 212 Bi with gradient concentration on donor cells for 6 h(C), 12h (D), 24 h (E), respectively (n=5). F, G, and H) The cytotoxic effect of supernatant from donor cells after 6 h (F), 12 h (G), and 24 h (H) 212 Bi-irradiation with gradient concentration on recipient cells for 24 h (n=5). I) Flow cytometry patterns of recipient cells and donor cells stained with annexin Ⅴ-FITC and propidium iodide (PI) for determining the apoptosis level. J) The cytotoxic effect of supernatant after 24 h 212 Bi-irradiation with gradient concentration on rotenone-pretreated recipient cells for 24 h.

Article Snippet: Calcein/Propidium Iodide (PI) cell viability/cytotoxicity assay kit, reduced glutathione (GSH) and oxidized glutathione disulfide (GSSG) assay kits, mitochondrial membrane potential assay kit with JC-1, and the Annexin V-Fluorescein isothiocyanate (FITC) apoptosis assay kit were obtained from Beyotime Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Concentration Assay, Irradiation, Flow Cytometry, Staining

Journal: Theranostics

Article Title: Role of damaged mitochondrial transfer in alpha-particle generator 212 Pb radiation-induced bystander effect

doi: 10.7150/thno.101922

Figure Lengend Snippet: The transfer of damaged mitochondria results in alpha-particle radiation-induced bystander effect. A) Schematic representation of the experimental design for the subsequent study. B, C, and D) The cytotoxic effect of various treatments with gradient concentration on recipient cells for 24 h was evaluated by CCK-8 assays (n=5 per group). E and F) Flow cytometry patterns of recipient cells stained with annexin Ⅴ-FITC and PI after various treatments for determining the apoptosis level. G) Cytotoxicity of various treatments on recipient cells by using co-staining with PI and Calcein-AM. H) Visualization of JC-1 monomer (Green) and JC-1 aggregate (Red) in recipient cells after various treatments.

Article Snippet: Calcein/Propidium Iodide (PI) cell viability/cytotoxicity assay kit, reduced glutathione (GSH) and oxidized glutathione disulfide (GSSG) assay kits, mitochondrial membrane potential assay kit with JC-1, and the Annexin V-Fluorescein isothiocyanate (FITC) apoptosis assay kit were obtained from Beyotime Biotechnology Co., Ltd. (Shanghai, China).

Techniques: Concentration Assay, CCK-8 Assay, Flow Cytometry, Staining