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: American Journal of Cancer Research

Article Title: YY1-induced lncRNA00511 promotes melanoma progression via the miR-150-5p/ADAM19 axis

doi:

Figure Lengend Snippet: LINC00511 was over-expressed in melanoma cells. A. The results from the GEO database showed that LINC00511 expression was significantly upregulated in melanoma tissues compared with normal tissues. The fold change (FC) of genes was assessed by log transformation. |log FC| > 2 and adjusted P < 0.05 were defined as the screened threshold. B. Based on the TCGA dataset, LINC00511 expression was found to be increased in melanoma tissues. C. The expression of LINC00511 in melanoma tissues and nor-tumor tissues by ISH (scale bars, 500 µm). D. Melanoma cells exhibited elevated levels of LINC00511 expression. E. Relative LINC00511 levels were in melanoma cells transfected with si-NC and si-LINC00511-1, -2. F, G. The proliferation of transfected melanoma cells was assessed using CCK8 and EdU experiments. H. The effect of transfection with si-NC, si-LINC00511-1, and -2 on the rate of apoptosis was evaluated using TUNEL analysis. ***, P ≤ 0.001; **, P ≤ 0.01; *, P ≤ 0.05; ns, not significant.

Article Snippet: Apoptosis analysis The TransDetect Annexin V-FITC/PI Apoptosis Detection Kit and TUNEL Apoptosis Assay Kit (Promega, Madison, WI, USA) were used to conduct apoptosis assays.

Techniques: Expressing, Transformation Assay, Transfection, TUNEL Assay

Journal: American Journal of Cancer Research

Article Title: YY1-induced lncRNA00511 promotes melanoma progression via the miR-150-5p/ADAM19 axis

doi:

Figure Lengend Snippet: Silencing ADAM19 suppressed melanoma progression in vitro and in vivo. A. The efficiency of si-ADAM19 knockdown was determined by qRT-PCR. B. The protein level of ADAM19 was illustrated by Western blot assay after knocking down ADAM19 in A375 and SK-Mel-28 cells. C, D. CCK8 and EdU experiments were used to evaluate the proliferation rate of transfected melanoma cells. E. The influence of ADAM19 knockdown on the rate of apoptosis was assessed by TUNEL analysis. F. Metastatic ability was assessed using the wound-healing assay. G. Relative expression levels of Bax, Bcl-2, MMP9, and PCNA were observed in A375 and SK-Mel-28 cells with ADAM19 knockdown. H. The tumor’s morphology, growth curve, volume, and weight were observed. I. The levels of ADAM19 in xenograft tissues were measured using qRT-PCR. J. The expression of Ki-67 was examined through IHC (scale bars, 200 µm). ***, P ≤ 0.001; **, P ≤ 0.01; *, P ≤ 0.05; ns, not significant.

Article Snippet: Apoptosis analysis The TransDetect Annexin V-FITC/PI Apoptosis Detection Kit and TUNEL Apoptosis Assay Kit (Promega, Madison, WI, USA) were used to conduct apoptosis assays.

Techniques: In Vitro, In Vivo, Knockdown, Quantitative RT-PCR, Western Blot, Transfection, TUNEL Assay, Wound Healing Assay, Expressing

Journal: American Journal of Cancer Research

Article Title: YY1-induced lncRNA00511 promotes melanoma progression via the miR-150-5p/ADAM19 axis

doi:

Figure Lengend Snippet: ADAM19 reversed the impact of miR-150-5p on melanoma cells. A. Expression levels of miR-150-5p were identified in melanoma cells after ADAM19 overexpression. B, C. Relative expression levels of ADAM19 are observed in A375 and SK-Mel-28 cells transfected with miR-150-5p. D, E. CCK8 and EdU assays to proliferation rate of transfected cells. F. The influence of ADAM19 and miR-150-5p mimics on the rate of apoptosis was evaluated through TUNEL analysis. G. Scratch wound healing assay is used to evaluate migration capacity. H. Relative expression levels of ADAM19, Bax, Bcl-2, MMP9, PCNA were observed in A375 and SK-Mel-28 cells transfected with ADAM19 and miR-150-5p mimics. ***, P ≤ 0.001; **, P ≤ 0.01; *, P ≤ 0.05; ns, not significant.

Article Snippet: Apoptosis analysis The TransDetect Annexin V-FITC/PI Apoptosis Detection Kit and TUNEL Apoptosis Assay Kit (Promega, Madison, WI, USA) were used to conduct apoptosis assays.

Techniques: Expressing, Over Expression, Transfection, TUNEL Assay, Wound Healing Assay, Migration

Journal: American Journal of Cancer Research

Article Title: YY1-induced lncRNA00511 promotes melanoma progression via the miR-150-5p/ADAM19 axis

doi:

Figure Lengend Snippet: LINC00511 acted as a sponge for miR-150-5p to up-regulate ADAM19 expression. A. The expression levels of ADAM19 in A375 and SK-Mel-28 cells transfected with si-LINC00511 and miR-150-5p inhibitor were identified using qRT-PCR. B. Relative expression levels of ADAM19, Bax, Bcl-2, MMP9, PCNA were observed in A375 and SK-Mel-28 cells transfected with si-LINC00511 and miR-150-5p inhibitor. C, D. CCK8 and EdU assays to proliferation rate of transfected cells. E. The influence of si-LINC00511 and miR-150-5p inhibitor on the rate of apoptosis was evaluated through TUNEL analysis. F. Metastasis ability was measured using wound-healing assay in melanoma cells transfected with si-LINC00511and miR-150-5p inhibitor. G. Gross appearance, tumor volume, and tumor weight of each group. H. IHC staining results (Ki-67). ***, P ≤ 0.001; **, P ≤ 0.01; *, P ≤ 0.05; ns, not significant.

Article Snippet: Apoptosis analysis The TransDetect Annexin V-FITC/PI Apoptosis Detection Kit and TUNEL Apoptosis Assay Kit (Promega, Madison, WI, USA) were used to conduct apoptosis assays.

Techniques: Expressing, Transfection, Quantitative RT-PCR, TUNEL Assay, Wound Healing Assay, Immunohistochemistry

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

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Paeoniflorin inhibits cell growth and induces cell cycle arrest through inhibition of FoxM1 in colorectal cancer cells.

doi: 10.1080/15384101.2017.1407892

Figure Lengend Snippet: Figure 1. PF inhibited cell proliferation and induced apoptosis and cell cycle arrest. A. Colon cancer cell viability was detected after PF treatment for 24 h, 48 h and 72 h, respectively, using MTT assay. P < 0.05, P < 0.01, compared to the control groups. B. Cell cycle analysis was performed by Flow cytometry in HCT116 cells treated with PF. C. PF-induced colon cancer cell apoptosis was accessed by Flow cytometry.

Article Snippet: Annexin apoptosis assay kit was purchased from Beyotime Biotechnology (Shanghai, China).

Techniques: MTT Assay, Control, Cell Cycle Assay, Flow Cytometry

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Paeoniflorin inhibits cell growth and induces cell cycle arrest through inhibition of FoxM1 in colorectal cancer cells.

doi: 10.1080/15384101.2017.1407892

Figure Lengend Snippet: Figure 3. Overexpression of FoxM1 abrogated PF-induced cell growth inhibition and apoptosis. A. Left panel: The expression of FoxM1 was detected by Western blotting analysis in HCT116 cells after FoxM1 overexpression plus PF treatment. Right panel; Quantitative results were illustrated for left panel. CTR: control; PF: Paeoniflorin; PF+cDNA: Paeoniflorin plus FoxM1 cDNA. B. Cell growth was measured by MTT assay after FoxM1 overexpression in combination with PF treatment. C. Cell cycle was accessed by Flow cytometry after FoxM1 overexpression in combination with PF treatment. D. Apoptotic cells were accessed by Flow cytometry after after FoxM1 overexpression in combination with PF treatment.

Article Snippet: Annexin apoptosis assay kit was purchased from Beyotime Biotechnology (Shanghai, China).

Techniques: Over Expression, Inhibition, Expressing, Western Blot, Control, MTT Assay, Flow Cytometry

Journal: Cell cycle (Georgetown, Tex.)

Article Title: Paeoniflorin inhibits cell growth and induces cell cycle arrest through inhibition of FoxM1 in colorectal cancer cells.

doi: 10.1080/15384101.2017.1407892

Figure Lengend Snippet: Figure 5. Knockdown of FoxM1 enhanced PF-mediated cell proliferation inhibition and apoptosis. A. The expression of FoxM1 was detected by Western blotting analysis in HCT116 cells after FoxM1 down-regulation plus PF treatment. Right panel: Quantitative results were illustrated for left panel. CTR: Control. PF: Paeoniflorin; PF+siRNA: Paeoniflorin plus FoxM1 siRNA. P < 0.05, vs control; # P < 0.05 vs FoxM1 siRNA transfection. B. MTT assay was performed to detect the effect of FoxM1 down-regulation in combination with PF treatment on HCT116 cell growth. P < 0.05, vs control; # P < 0.05 vs FoxM1 siRNA transfection. C. Cell cycle was accessed by Flow cytometry after FoxM1 downregulation in combination with PF treatment. D. Apoptotic cells were detected by Flow cytometry in HCT116 cells after FoxM1 siRNA transfection and PF treatment.

Article Snippet: Annexin apoptosis assay kit was purchased from Beyotime Biotechnology (Shanghai, China).

Techniques: Knockdown, Inhibition, Expressing, Western Blot, Control, Transfection, MTT Assay, Flow Cytometry

Journal: International Journal of Oncology

Article Title: Eupalinolide A induces autophagy via the ROS/ERK signaling pathway in hepatocellular carcinoma cells in vitro and in vivo

doi: 10.3892/ijo.2022.5421

Figure Lengend Snippet: Effect of EA on apoptosis and necroptosis of human hepatocellular carcinoma cells. (A and B) Western blot analysis of the expression levels of apoptosis and necroptosis-related proteins at 48 h in MHCC97-L and HCCLM3 cells. β-actin was used as a control. (C and D) Effect of EA on apoptosis in MHCC97-L and HCCLM3 cells, as determined by flow cytometry. n=5. (E) Viability of MHCC97-L and HCCLM3 cells treated with or without apoptosis or necroptosis inhibitors was measured by Cell Counting Kit 8 assays. n=6. All data are shown as the mean ± SD. *** P<0.001. EA, eupalinolide A; C-, cleaved; p-, phosphorylated.

Article Snippet: The cells were then collected, washed with cold PBS and suspended in 200 μ l binding buffer (Annexin V-FITC/PI apoptosis assay kit; Neobioscience Technology Co., Ltd.) at a density of 5×10 5 /ml.

Techniques: Western Blot, Expressing, Control, Flow Cytometry, Cell Counting