Journal: Molecular Biology of the Cell

Article Title: Image-based drug screen identifies HDAC inhibitors as novel Golgi disruptors synergizing with JQ1

doi: 10.1091/mbc.E17-03-0176

Figure Lengend Snippet: Image-based chemical compound screen identifies HDAC inhibitors and DNA-damaging agents as novel Golgi-dispersing compounds. To identify novel compounds that modulate Golgi morphology, a screening platform was established. (A) Screening pipeline including cell seeding (A549 cells), treatment with compound library, staining for the cis -Golgi (GM130), the nucleus (Hoechst), the ER stress marker (GRP78), cytoplasm (Phalloidin), and image acquisition and processing. (B) Representative images of the negative control (vehicle-treated cells) as well as positive controls (BFA, doxorubicin, and nocodazole) and newly discovered Golgi-fragmenting drugs (Bleomycin, Vorinostat, 4-iodo-SAHA, Trichostatin A, Givinostat, and Pracinostat) are displayed. (C) Following image analysis, to exclude potential plate effects, the Golgi area of cells treated with the chemical library was normalized to the vehicle-treated sample present within the same plate. The corresponding survival ratios of treated cells are also shown. (D) Detailed view of compounds are shown, of which at least two of three replicates caused a ≥1.5-fold increase in Golgi area. The compound panel includes positive controls and novel Golgi-fragmenting compounds, which were selected for further investigation. *** p < 0.001 vs. #; see Materials and Methods .

Article Snippet: Compounds were obtained from the following companies: brefeldin A (Sigma-Aldrich), golgicide A (Santa Cruz Biotechnology), monensin (Enzo Life Sciences), AG-1478 (Sigma), tunicamycin (Santa Cruz Biotechnology), thapsigargin (Santa Cruz Biotechnology), nocodazole (Santa Cruz Biotechnology), (+)-JQ1 (Cayman Chemical), CBP30 (TargetMol), doxorubicin (Sigma), etoposide (Sigma), teniposide (Santa Cruz Biotechnology), mitomycin-C (Santa Cruz Biotechnology), cisplatin (Santa Cruz Biotechnology), hydroxyurea (Sigma), 5-fluorouracil (Sigma), gemcitabine (Santa Cruz Biotechnology), irinotecan (Santa Cruz Biotechnology), bleomycin (Santa Cruz Biotechnology), NU7441 (Selleckchem), KU55933 (Sigma), Flavopiridol (Santa Cruz Biotechnology), phorbol 12-myristate 13-acetate (PMA; Santa Cruz Biotechnology), Panobinostat (Selleckchem), Tubastatin (Selleckchem), Entinostat (Santa Cruz Biotechnology), Pracinostat (Selleckchem), Givinostat (Selleckchem), Triptolide (Santa Cruz Biotechnology), α-Amanitin (Santa Cruz Biotechnology), and Z-VAD-FMK (Santa Cruz Biotechnology).

Techniques: Drug discovery, Staining, Marker, Negative Control

Journal: Molecules (Basel, Switzerland)

Article Title: Cytostatic Bacterial Metabolites Interfere with 5-Fluorouracil, Doxorubicin and Paclitaxel Efficiency in 4T1 Breast Cancer Cells.

doi: 10.3390/molecules29133073

Figure Lengend Snippet: Figure 9. Cadaverine and indolepropionic acid interfere with the cytostatic effect of doxorubicin. 4T1 cells were plated in 96-well plates (1500 cells/well). Cells were treated with doxorubicin alone or in combination with CAD (0.8 µM), IS (4 µM) or IPA (1 µM) for 48 h, and then cell numbers were determined by MTT assay. Data are presented as means ± SEM, from at least three biological replicates. Individual assays were measured in quadruplicate or in triplicate. Values were normalized to vehicle-treated cells (absorbance is equal to 1). Nonlinear regression (Graphpad “[Inhibitor] vs. response (four parameters)” utility) was performed on datasets to obtain IC50 and Hill slope values. Normality was determined for the inhibitory curves using the D’Agostino and Pearson normality test, while for the IC50 values and the Hill slope values the Shapiro–Wilk test was used. Statistical difference between the inhibitory curves was determined using a two-way ANOVA test, and all data points were compared with each other (in Tukey post hoc tests). For the comparison of the IC50 and Hill slope values, a non-paired, two-sided t-test was applied. ### indicates p < 0.001 for DOX-treated vs. vehicle-treated cells. * represents significance at p < 0.05 between the indicated groups. Abbreviations: CAD—cadaverine; DOX—doxorubicin; IPA—indolepropionic acid; and IS—indoxylsulfate.

Article Snippet: Liposomal Encapsuled Doxorubicin (DOX-NP, cat # 300112) was purchased from Avanti Polar Lipids (Alabaster, AL, USA) and a stock solution of 50 mM was prepared.

Techniques: MTT Assay, Comparison

Journal: Molecules

Article Title: Exploring the Interplay between Drug Release and Targeting of Lipid-Like Polymer Nanoparticles Loaded with Doxorubicin

doi: 10.3390/molecules26040831

Figure Lengend Snippet: Figure 1. Illustration of the synthesis and adsorption process involved in drug delivery using lipid-like polylactide-co- glycolide (PLGA) nanoparticles loaded with doxorubicin.

Article Snippet: Plasma doxorubicin concentrations were calculated from two calibration curves using the chemical reference standard (CRS) of doxorubicin (EP CRS, 99.0%, EDQM).

Techniques: Adsorption

Journal: Oncotarget

Article Title: Tumor targeting with pH-responsive poly(2-oxazoline)-based nanogels for metronomic doxorubicin treatment

doi: 10.18632/oncotarget.24806

Figure Lengend Snippet: Lysosomal cellular compartments were stained green using LysoTracker Green DND-26 and the nucleus was labeled with Hoechst 33342 (blue). The fluorescence of DOX is depicted in red and the Alexafluor label of the polymer is shown in white.

Article Snippet: Live cell CLSM images were acquired using a Zeiss LSM 880, Elyra PS.1 system (Carl Zeiss, Germany) with excitation wavelengths/emission filters of 405nm/BP 405–480 nm for Hoechst 33342, 488 nm/BP 505 to 530 nm for LysoTracker ® Green DND-26 and 488 nm/BP 585 to 615 nm for DOX and 633 nm/BP 724 to 777 nm for Alexafluor 660 ® .

Techniques: Staining, Labeling, Fluorescence