Journal: Oncotarget

Article Title: Evaluation of in vitro effects of various targeted drugs on plasma cells and putative neoplastic stem cells in patients with multiple myeloma

doi: 10.18632/oncotarget.11593

Figure Lengend Snippet: Effects of various targeted drugs on proliferation of myeloma cell lines

Article Snippet: A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA).

Techniques: Produced

Journal: Oncotarget

Article Title: Evaluation of in vitro effects of various targeted drugs on plasma cells and putative neoplastic stem cells in patients with multiple myeloma

doi: 10.18632/oncotarget.11593

Figure Lengend Snippet: Effects of the most effective targeted drugs on proliferation of primary neoplastic BM cells

Article Snippet: A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA).

Techniques: Produced

Journal: Oncotarget

Article Title: Evaluation of in vitro effects of various targeted drugs on plasma cells and putative neoplastic stem cells in patients with multiple myeloma

doi: 10.18632/oncotarget.11593

Figure Lengend Snippet: Effects of various targeted drugs on survival (apoptosis) of myeloma cell lines

Article Snippet: A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA).

Techniques: Produced

Journal: Oncotarget

Article Title: Evaluation of in vitro effects of various targeted drugs on plasma cells and putative neoplastic stem cells in patients with multiple myeloma

doi: 10.18632/oncotarget.11593

Figure Lengend Snippet: MM cell lines (MM.1S, NCI-H929, OPM-2, RPMI-8226, U-266) were incubated in control medium (Co) or in various concentrations of 17AAG, BI2536, or BEZ235 (0.001-1 μM) at 37°C for 48 hours. Then, the percentage of apoptotic cells was determined by AnnexinV/PI staining and flow cytometry. Results show the percentage of AnnexinV/PI+ cells and represent the mean±S.D. from 3 independent experiments. Asterisk (*): p<0.05.

Article Snippet: A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA).

Techniques: Incubation, Control, Staining, Flow Cytometry

Journal: Oncotarget

Article Title: Evaluation of in vitro effects of various targeted drugs on plasma cells and putative neoplastic stem cells in patients with multiple myeloma

doi: 10.18632/oncotarget.11593

Figure Lengend Snippet: Primary BM cells derived from 6 patients with MM were incubated in control medium (Co) or in medium containing 17AAG, BI2536, or BEZ235 (each 1 μM) at 37°C for 48 hours. Thereafter, cells were stained with antibodies against AnnexinV A. or active caspase-3 B. by multicolor flow cytometry as described in the text. The following subsets of cells were examined: CD138 + MM cells (black bars), CD138 − /CD27 + /CD20 + putative MMSC (grey bars), CD34 + /CD38 − hematopoietic stem cells (open bars), and CD34 + /CD38 + cells (hatched bars). Results are expressed as percent AnnexinV+ cells (A) or percent active caspase-3+ cells (B) and represent the mean±S.D. from 6 independent experiments. Asterisk (*): p<0.05.

Article Snippet: A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA).

Techniques: Derivative Assay, Incubation, Control, Staining, Flow Cytometry

Journal: Oncotarget

Article Title: Evaluation of in vitro effects of various targeted drugs on plasma cells and putative neoplastic stem cells in patients with multiple myeloma

doi: 10.18632/oncotarget.11593

Figure Lengend Snippet: MM.1S cells (upper left panel), NCI-H929 cells (upper right panel), OPM-2 cells (middle left panel), RPMI-8226 cells (middle right panel) and U-266 cells (lower panel) were incubated in control medium (Co) or various concentrations of 17AAG, BI2536, or BEZ235 (0.001-1 μM each) at 37°C for 48 hours. Then, cell cycle distribution was analyzed by flow cytometry as described in the text. Asterisk (*): p<0.05.

Article Snippet: A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA).

Techniques: Incubation, Control, Flow Cytometry

Journal: Oncotarget

Article Title: Evaluation of in vitro effects of various targeted drugs on plasma cells and putative neoplastic stem cells in patients with multiple myeloma

doi: 10.18632/oncotarget.11593

Figure Lengend Snippet: MM.1S cells A. , OPM-2 cells B. , and RPMI-8226 cells C. were incubated in control medium (Co), in medium containing individual drugs alone, or in medium containing drug combinations (at fixed ratio) at 37°C for 48 hours. Then, uptake of 3 H-thymidine was measured. MM.1S cells (A) were incubated in various concentrations of BI2536 ( ○ - ○ ), BEZ235 ( ◇ - ◇ ), or combinations of both drugs ( ◾ - ◾ ). OPM-2 cells (B) were incubated with various concentrations of BI2536 ( ○ - ○ ) or obatoclax ( ◇ - ◇ ) or combinations of both drugs ( ○ - ○ ). RPMI-8226 cells (C) were incubated with various concentrations of 17AAG ( ○ - ○ ) or BEZ235 ( ◇ - ◇ ) or combinations of both drugs ( ○ - ○ ). Results show the percentage of 3 H-thymidine uptake compared to medium control and represent the mean±S.D. of one typical experiment.

Article Snippet: A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA).

Techniques: Incubation, Control

Journal: Oncotarget

Article Title: Evaluation of in vitro effects of various targeted drugs on plasma cells and putative neoplastic stem cells in patients with multiple myeloma

doi: 10.18632/oncotarget.11593

Figure Lengend Snippet: Effects of various drug combinations on proliferation of myeloma cell lines

Article Snippet: A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA).

Techniques: Produced

Journal: mBio

Article Title: Genetic Analysis of Candida auris Implicates Hsp90 in Morphogenesis and Azole Tolerance and Cdr1 in Azole Resistance

doi: 10.1128/mBio.02529-18

Figure Lengend Snippet: Perturbation of HSP90 results in a loss of viability of C. auris but does not affect azole resistance. (A) Spotting of C. auris wild-type and tetO-HSP90 strains on YPD or YPD agar plus doxycycline (DOX) plates (right panel). C. albicans wild-type and tetO-HSP90 / tetO-HSP90 strains were included for comparison (left panel). A high concentration (50 μg/ml) of DOX was used to ensure strong repression of HSP90 expression. Overnight cultures were diluted 1,000-fold, at which point 5 μl was spotted in 100-fold dilutions. Plates were incubated at 30°C for 48 h. (B) Fluconazole Etest strip in the presence and absence of DOX. A total of 1 × 10 6 cells of wild-type and tetO- repressible HSP90 strains were plated on YPD agar plates with fluconazole Etest strips in the absence and presence of DOX (0.1 μg/ml or 10 μg/ml). The plates were incubated at 30°C for 48 h. (C) Checkerboard assays with geldanamycin and fluconazole. C. albicans clinical isolate CaCi2 and C. auris isolate Ci6684 were inoculated with a 2-fold gradient of geldanamycin in combination with a 2-fold gradient of fluconazole. Cultures were incubated at 30°C for 48 h. Heat maps represent relative growth levels determined from averages of results of technical replicates normalized to the data from a no-drug well.

Article Snippet: Approximately 1 × 10 3 cells were inoculated with a 2-fold gradient matrix of fluconazole (Carbosynth) or of geldanamycin (LC Laboratories) or of a combination of the two, as indicated, in 96-well microtiter plates to reach the final volume of 200 μl in YPD.

Techniques: Concentration Assay, Expressing, Incubation, Stripping Membranes

Journal: mBio

Article Title: Genetic Analysis of Candida auris Implicates Hsp90 in Morphogenesis and Azole Tolerance and Cdr1 in Azole Resistance

doi: 10.1128/mBio.02529-18

Figure Lengend Snippet: Hsp90 mediates tolerance of fluconazole in C. auris . (A) MIC assay for fluconazole in a panel of C. auris clinical isolates. Data were analyzed as described for Fig. 1C . (B) Checkerboard assays with geldanamycin and fluconazole. C. auris strains Ci6684, CDC-382, CDC-387, and CDC-388 were inoculated with a 2-fold gradient of geldanamycin in combination with a 2-fold gradient of fluconazole. Data were analyzed as described for Fig. 1C . After measurement of the growth, cultures were spotted onto drug-free YPD agar plates and allowed to grow at 30°C for 24 h to check for cidality.

Article Snippet: Approximately 1 × 10 3 cells were inoculated with a 2-fold gradient matrix of fluconazole (Carbosynth) or of geldanamycin (LC Laboratories) or of a combination of the two, as indicated, in 96-well microtiter plates to reach the final volume of 200 μl in YPD.

Techniques:

Journal: mBio

Article Title: Genetic Analysis of Candida auris Implicates Hsp90 in Morphogenesis and Azole Tolerance and Cdr1 in Azole Resistance

doi: 10.1128/mBio.02529-18

Figure Lengend Snippet: C. auris undergoes filamentous growth under conditions of compromised Hsp90 function. (A) Overnight cultures of C. albicans (strain SN95) and C. auris (strain Ci6684) or of the respective strains with HSP90 under the control of a tetO -repressible promoter were subcultured in YPD without or with doxycycline. Images were captured after 6 h postsubculture. (B) Overnight cultures of wild-type C. albicans and C. auris strains were subcultured in YPD without or with geldanamycin (10 μM for C. albicans and 80 μM for C. auris ) or hydroxyurea (50 mM). Images were captured after 6 h of drug treatment. (C) Overnight cultures of C. albicans and C. auris were subcultured in canonical C. albicans filament-inducing cues. These include 37°C plus 10% serum (Serum), 37°C plus Spider medium (Spider), 37°C RPMI medium, and 42°C. Microscopy images were acquired after 6 h of incubation. The scale bars represent 20 μm in all panels.

Article Snippet: Approximately 1 × 10 3 cells were inoculated with a 2-fold gradient matrix of fluconazole (Carbosynth) or of geldanamycin (LC Laboratories) or of a combination of the two, as indicated, in 96-well microtiter plates to reach the final volume of 200 μl in YPD.

Techniques: Microscopy, Incubation

Journal: mBio

Article Title: Genetic Analysis of Candida auris Implicates Hsp90 in Morphogenesis and Azole Tolerance and Cdr1 in Azole Resistance

doi: 10.1128/mBio.02529-18

Figure Lengend Snippet: CUG clade species undergo filamentous growth under conditions of compromised Hsp90 function. Overnight cultures of C. albicans , C. dubliniensis , C. tropicalis , L. elongisporus , C. lusitaniae , C. auris , S. cerevisiae , and C. glabrata were subcultured in YPD without or with geldanamycin (GdA). (Top left) A phylogenetic tree of the yeast species was constructed using 1,570 single-copy protein-coding genes, maximum likelihood inference based on 1,000 replicates, and RAxML v7.7.8. Scale bar, mean number of nucleotide substitutions per site. (Top right) Microscopy images were acquired after 6 h of incubation. Scale bar, 20 μm. The concentration of geldanamycin used for each species is indicated on the micrograph. (Bottom) Proportions of yeast to filament were quantified.

Article Snippet: Approximately 1 × 10 3 cells were inoculated with a 2-fold gradient matrix of fluconazole (Carbosynth) or of geldanamycin (LC Laboratories) or of a combination of the two, as indicated, in 96-well microtiter plates to reach the final volume of 200 μl in YPD.

Techniques: Construct, Microscopy, Incubation, Concentration Assay