Journal: The Journal of investigative dermatology

Article Title: Chloroquine Promotes Apoptosis in Melanoma Cells by Inhibiting BH3 domain Mediated PUMA Degradation

doi: 10.1038/jid.2013.56

Figure Lengend Snippet: a & b) Indicated melanoma cells were exposed to etoposide or different concentrations of CQ and the cell extracts were blotted with PUMA, p53, p21 and actin antibodies. c) Shows quantification of PUMA signal intensity from a & b. d & e) Melanoma cells were treated with etoposide or CQ and the relative levels of PUMA transcripts were assayed by q-PCR.

Article Snippet: Bafilomycin A, chloroquine diphosphate, leupeptin, and MG-132 were obtained from Sigma, ALLN, etoposide, and Z-Vad-Fmk from R&D Systems (Minneapolis), E64d and Pepstatin A (Cayman Chemical) and lactacystin (AG Scientific).

Techniques:

Journal: MedComm – Oncology

Article Title: Gene LY96 is an M2 macrophage‐related biomarker and is associated with immunosuppression in renal cell carcinoma

doi: 10.1002/mog2.52

Figure Lengend Snippet: FIGURE 6 Immunotherapy efficacy prediction and drug prediction for lymphocyte antigen 96 (LY96). (A) LY96 is positively associated with immune scores. (B–D) Comparison of LY96 expression level between ICI responder and nonresponder. (E) Twenty‐seven drugs are predicted by TAiC database for targeting LY96. (F) Seven drugs are predicted by Cellminer database. (G) Vemurafenib and etoposide are the intersections of two predictions. (H, I) Molecular docking of LY96 and two drugs. False discovery rate (FDR) or p value < 0.05 is defined as significant. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05.

Article Snippet: Etoposide (Selleck) at different concentrations (0, 1, 2, 3, 4, 5 μM) was added to the culture medium and cocultured with M2 macrophages for 24 h. Afterward, the cells were collected for further analysis.

Techniques: Comparison, Expressing

Journal: Scientific Reports

Article Title: Transactivation of human osteoprotegerin promoter by GATA-3

doi: 10.1038/srep12479

Figure Lengend Snippet: ( A ) HEK cells expressing either a scrambled shRNA or a GATA-3 shRNA expressing plasmid were treated with etoposide (10 μM), and apoptosis was assayed by Western blot (left). HEK cells expressing a GATA-3 shRNA expressing plasmid were treated with purified OPG protein for 2 h before treatment with etoposide, and apoptosis was assayed by Western blot (right). Full-length blots are presented in . ( B ) HEK cells expressing either a control plasmid or a GATA-3 shRNA expressing plasmid were treated with TNF-α (2 μg/mL) with or without pre-treatment with OPG for 2 h. Subsequent cell death was measured using TUNEL assays (left), and quantified with bar graphs (right). *p < 0.05, n = 3.

Article Snippet: The apoptosis inducing agents, tumor necrosis factor alpha (TNF-α) (#8902) and etoposide (#341205), were from Cell Signaling and EMD Millipore (USA), respectively.

Techniques: Expressing, shRNA, Plasmid Preparation, Western Blot, Purification, Control, TUNEL Assay

Journal: Plants

Article Title: The Role of DNA Topoisomerase Binding Protein 1 (TopBP1) in Genome Stability in Arabidopsis

doi: 10.3390/plants10122568

Figure Lengend Snippet: Defects in DSB repair and mitosis in topbp1 mutant. ( a ) Diagram with the mean number of leaves per seedling in the WT and topbp1 mutant grown just in MS medium or supplemented with cisplatin (30 μM) or cisplatin + etoposide (5 μM). Data collected from three independent experiments ( n = 100 per treatment and day). ( b ) Mitotic anaphases of the WT and topbp1 . Statistical differences between the WT and topbp1 for each treatment analysed by Mann–Whitney test, *** p < 0.001; ns = not significant. Comparisons among treatments within the same genetic background are shown in . Scale bar is 5 μm.

Article Snippet: Etoposide (Tocris Bioscience, Bristol, UK) at 5 μM was also used in conjunction with the DSB repair assessment.

Techniques: Mutagenesis, MANN-WHITNEY

Journal: Plants

Article Title: The Role of DNA Topoisomerase Binding Protein 1 (TopBP1) in Genome Stability in Arabidopsis

doi: 10.3390/plants10122568

Figure Lengend Snippet: Results of the pairwise comparison of the mean number of leaves per seedling untreated (MS), treated with cisplatin, and treated with cisplatin + etoposide (Cis + Etop) by the Kruskal–Wallis test followed by Dunn’s post-hoc test in the WT and topbp1 at days 7, 12, and 16.

Article Snippet: Etoposide (Tocris Bioscience, Bristol, UK) at 5 μM was also used in conjunction with the DSB repair assessment.

Techniques: Comparison

Journal: Plants

Article Title: The Role of DNA Topoisomerase Binding Protein 1 (TopBP1) in Genome Stability in Arabidopsis

doi: 10.3390/plants10122568

Figure Lengend Snippet: Meiotic stages of WT plants treated with different topoisomerase II inhibitors. ( a ) Plants were treated with TOPII inhibitors in two ways: (i) a 2 h pulse (P) or (ii) continuous (C). In both cases, flower buds were fixed at 12 h, 28 h, or 38 h after treatment. ( b – m ) Images of pollen mother cells at different stages of meiosis of the WT treated with TOPII inhibitors. ( b ) Anaphase I treated with merbarone 1 μM (P) fixed at 38 h showing an anaphase bridge. ( c ) Anaphase I treated with merbarone 1 μM (C) fixed at 38 h showing a chromosome fragment. ( d ) Anaphase II treated with merbarone 10 μM (P) fixed at 38 h showing chromosome mis-segregation. ( e ) Telophase II treated with merbarone 1 μM (C) fixed at 38 h showing micronuclei. ( f ) Anaphase I treated with etoposide 0.05 μM (P) fixed at 38 h showing a broken anaphase bridge. ( g ) Anaphase II treated with etoposide 0.05 μM (C) fixed at 38 h showing chromosome mis-segregation. ( h ) Telophase II treated with etoposide 0.05 μM (P) fixed at 28 h showing a micronucleus. ( i ) Telophase II treated with etoposide 5 μM (P) fixed at 28 h showing a micronucleus. ( j ) Anaphase I treated with ICRF-187 0.1 μg/mL (P) fixed at 38 h showing an anaphase bridge. ( k ) Anaphase I treated with ICRF-187 100 μg/mL (C) fixed at 28 h showing two anaphase bridges. ( l ) Metaphase II/anaphase II treated with ICRF-187 100 μg/mL (C) fixed at 28 h showing an anaphase bridge. ( m ) Telophase II treated with ICRF-187 0.1 μg/mL (P) fixed at 38 h showing a micronucleus. Arrows indicate errors in meiotic divisions. Scale bar 10 μm.

Article Snippet: Etoposide (Tocris Bioscience, Bristol, UK) at 5 μM was also used in conjunction with the DSB repair assessment.

Techniques:

Journal: Oncology reports

Article Title: Zoledronic acid inhibits proliferation of human fibrosarcoma cells with induction of apoptosis, and shows combined effects with other anticancer agents.

doi: 10.3892/or_00000851

Figure Lengend Snippet: Figure 5. Growth inhibitory effects of concurrent treatment with ZOL and anticancer agents on human fibrosarcoma cell lines. The capacity of ZOL and several antitumor agents to inhibit the growth of HT1080 cells was determined by employing the trypan blue dye exclusion method. Data from three independent experiments were collected, and Student's t-test was used to evaluate the efficacy of concurrent treatment with ZOL and other agents and to compare the effects of each anticancer agent alone. P-values of <0.05 were considered statistically significant and derived from two-sided statistical tests. X-axis: a, control; b, 1.2 μM of ZOL alone; c, 0.5xIC50 of antitumor drug alone; d, combination of b with c; e, 1.0xIC50 of antitumor drug; f, combination of b with d. Y-axis is cell counts (x105). (A) doxorubicin; (B) cisplatin; (C) etoposide; (D) 5-fluorouracil; (E) docetaxel; (F) paclitaxel; (G) gemcitabine; (H) methotrexate.

Article Snippet: Doxorubicin (from Toronto Research Chemicals, Inc., Toronto, Canada), 5-fluorouracil (from Nacalai Tesque, Inc., Kyoto, Japan), cisplatin, paclitaxel, docetaxel, gemcitabine (from LKT laboratories, Inc., St. Paul, MN, USA), etoposide (from CalbiochemNovabiochem, Cor(Merck KGaA), Darmstadt, Germany), and methotrexate (from Sigma Aldrich, Tokyo, Japan) were purchased from commercial sources.

Techniques: Derivative Assay, Control

Journal: Molecular cancer research : MCR

Article Title: Heat Shock Protein 70 (Hsp70) Suppresses RIP1-Dependent Apoptotic and Necroptotic Cascades

doi: 10.1158/1541-7786.MCR-17-0408

Figure Lengend Snippet: (A) Overexpression of Bcl-xL does not block JG-98 cytotoxicity. MDA-MB-231 cells were treated for 24 hrs with JG-98 (10 μM), 17-DMAG (10 μM), bortezomib (40 nM) or etoposide (20 μM). MTT results are the mean average of triplicates and error bars represent SEM. *p value < 0.05. (B) Jurkat cells over-expressing Bcl-xL were treated with indicated compounds for 24 hours. Viability was determined by trypan blue staining. Results are the average of two experiments performed in triplicate. Error is SEM. **p value < 0.01. (C) Bcl-xL overexpression provides partial resistance to compounds, except JG-98. MDA-MB-231 cells were treated for 72 hours and cell growth determined by MTT assays. Results are the average of two independent experiments performed in triplicate. Error is SEM.

Article Snippet: The following reagents were purchased from Sigma-Aldrich: Necrostatin-1, Bortezomib; Enzo: z-VAD.fmk; Millipore: Necrosulfonamide; LC Labs: 17-DMAG; StressMarq: VER-155008; and Teva Pharmaceuticals: Etoposide.

Techniques: Over Expression, Blocking Assay, Expressing, Staining

Journal: Molecular cancer research : MCR

Article Title: Heat Shock Protein 70 (Hsp70) Suppresses RIP1-Dependent Apoptotic and Necroptotic Cascades

doi: 10.1158/1541-7786.MCR-17-0408

Figure Lengend Snippet: (A) RIP1 KO Jurkat cells are resistant to JG-98. Viability was determined by trypan blue exclusion. Cells were treated for 24 hrs with JG-98 (10 μM), 17-DMAG (10 μM), bortezomib (40 nM), etoposide (20 μM). Results are the average of three independent experiments performed in triplicate. ns = not significant; * p < 0.05. (B) JG-98 activity requires RIP1 kinase. MDA-MB-231 cells were pretreated with 20 μM necrostatin-1 for 1 hour prior to addition of compounds. Viability was determined by three independent MTT assays performed in quintuplicate. Error is SEM. ns = not significant; ***p < 0.0001. (C) JG-98 induces degradation of RIP1 modulators. MDA-MB-231 cells were treated for 24 hours. Results represent experiments performed in triplicate. (D) RIP1 regulators are rapidly degraded in response to JG-98. MDA-MB-231 cells were treated with JG-98 (10 μM). Results are representative of duplicates.

Article Snippet: The following reagents were purchased from Sigma-Aldrich: Necrostatin-1, Bortezomib; Enzo: z-VAD.fmk; Millipore: Necrosulfonamide; LC Labs: 17-DMAG; StressMarq: VER-155008; and Teva Pharmaceuticals: Etoposide.

Techniques: Activity Assay