Journal: Journal of Medicinal Chemistry

Article Title: Doxorubicin and Aclarubicin: Shuffling Anthracycline Glycans for Improved Anticancer Agents

doi: 10.1021/acs.jmedchem.0c01191

Figure Lengend Snippet: Figure 2. Evaluation of DNA break capacity and histone evicting activity of hybrid structures 2−11 and parent compounds doxorubicin (1) and aclarubicin (12). (A) K562 cells were treated for 2 h with 10 μM of the indicated drugs, etoposide was used as a positive control for DNA double-strand breaks. γH2AX levels were examined by Western blot. Actin was used as a loading control, and molecular weight markers are as indicated. (B) Quantification of the γH2AX signal normalized to actin. Results are presented as mean ± standard deviation (SD) of three independent experiments. Ordinary one-way analysis of variance (ANOVA) with Dunnett’s multiple comparison test; ns, not significant; ****P < 0.0001. (C) Quantification of broken DNA relative to intact DNA as analyzed by constant-field gel electrophoresis (CFGE). Etoposide was used as a positive control for DNA double-strand breaks. Results are presented as mean ± SD of three independent experiments. Ordinary one-way ANOVA with Dunnett’s multiple comparison test; *P < 0.05, ****P < 0.0001 is indicated, all others are not significant. (D) Quantification of the release of fluorescent PAGFP-H2A from the photoactivated nuclear regions after administration of 10 μM of the indicated drugs. Results are shown as mean ± SD of 10−20 cells from at least three independent experiments. Ordinary two-way ANOVA with Dunnett’s multiple comparison test; ns, not significant; ****P < 0.0001. See also Figures S1 and S2.

Article Snippet: Doxorubicin was obtained from Accord Healthcare Limited, U.K., aclarubicin from Santa Cruz Biotech, and etoposide from Pharmachemie, Haarlem, The Netherlands.

Techniques: Activity Assay, Positive Control, Western Blot, Control, Molecular Weight, Standard Deviation, Comparison, Nucleic Acid Electrophoresis

Journal: Journal of Medicinal Chemistry

Article Title: Doxorubicin and Aclarubicin: Shuffling Anthracycline Glycans for Improved Anticancer Agents

doi: 10.1021/acs.jmedchem.0c01191

Figure Lengend Snippet: Figure 3. Cytotoxicity of compounds 1−12. (A, B) K562 cells were treated for 2 h at the indicated doses (higher doses in (A), lower doses in (B)) of the various hybrid compounds followed by drug removal. Cell survival in MelJuSo (C), human colorectal carcinoma cell line HCT116 (D), human prostate tumor cell line PC3 (E) and DU145 (F), and human glioblastoma cell line U87 (G). Cells were treated for 2 h at indicated dose followed by drug removal. Cell viability was measured by a Cell-Titerblue assay 72 h post-treatment. Data are shown as mean ± SD from three different experiments. (H) Table showing the IC50 values for the different doxorubicin/aclarubicin hybrid compounds for the indicated cell lines. See also Figure S3A, Supporting Information.

Article Snippet: Doxorubicin was obtained from Accord Healthcare Limited, U.K., aclarubicin from Santa Cruz Biotech, and etoposide from Pharmachemie, Haarlem, The Netherlands.

Techniques:

Journal: Osteoarthritis and Cartilage Open

Article Title: Iron triggers the early stages of cartilage degeneration in vitro: The role of articular chondrocytes

doi: 10.1016/j.ocarto.2021.100145

Figure Lengend Snippet: Fig. 3. Iron loading induces morphological alter- ations in chondrocytes. Transmission electron micro- scopy images illustrate the ultrastructure of chondrocytes in freshly collected cartilage (A) or in cartilage cultured for 9 days with sodium citrate (control) (B) or ferric citrate (iron loading condition) (C–F). Whilst the morphology of control chondrocytes was indistinct from that of cells from freshly collected cartilage, iron-treated chondrocytes presented side- rosomes (white arrows) and a number of morpho- logical changes consistent with cell death by chondroptosis. (D) is a magnification of the area within the square in (C), showing a siderosome and abundant dilated cisternae of rough ER. A chon- droptotic cell with convoluted nucleus with condensed chromatin, abundant Golgi and vacuoles (arrow heads), and extrusion of cellular material into the extracellular space (black arrows) is represented in (E). An empty lacuna is depicted in (F), where the remnants of a dead chondrocyte are identified by a dotted line. Samples were contrasted with uranile acetate and lead citrate. Original magnification: 15000 . Bar¼1 μm.

Article Snippet: Cartilage was also incubated with ER stress/ chondroptosis inducers (tunicamycin or thapsigargin, Santa Cruz Biotechnology) or with ferroptosis inducers (erastin; RSL-3; buthioninesulfoximine, BSO) for 9 days.

Techniques: Transmission Assay, Cell Culture, Control

Journal: Oncogene

Article Title: Degradation of the E7 human papillomavirus oncoprotein by the ubiquitin-proteasome system: targeting via ubiquitination of the N-terminal residue.

doi: 10.1038/sj.onc.1203989

Figure Lengend Snippet: Figure 2 Degradation of E7 in cells is sensitive to proteasome inhibition. Cos 7 cells were transiently transfected with cDNA coding for E7. Degradation of the protein was monitored in a pulse-chase labeling and immunoprecipitation experiment in the absence or presence of the cell permeable proteasome inhibitor clasto-lactacystin b-lactone as described under Materials and methods

Article Snippet: Antibody against E7 was from Santa Cruz. clasto-lactacystin b-lactone was from Calbiochem.

Techniques: Inhibition, Transfection, Pulse Chase, Labeling, Immunoprecipitation

Journal: Oncogene

Article Title: Degradation of the E7 human papillomavirus oncoprotein by the ubiquitin-proteasome system: targeting via ubiquitination of the N-terminal residue.

doi: 10.1038/sj.onc.1203989

Figure Lengend Snippet: Figure 4 Degradation of lysine-less E7 in cells is sensitive to proteasome inhibition. Cos 7 cells were transiently transfected with cDNA coding for lysine-less E7. Degradation of the protein was monitored in a pulse-chase labeling and immunoprecipitation experiment in the absence or presence of the cell permeable proteasome inhibitor clasto-lactacystin b-lactone as described under Materials and methods

Article Snippet: Antibody against E7 was from Santa Cruz. clasto-lactacystin b-lactone was from Calbiochem.

Techniques: Inhibition, Transfection, Pulse Chase, Labeling, Immunoprecipitation