Journal: Antibiotics

Article Title: Antifungal Drug Repurposing

doi: 10.3390/antibiotics9110812

Figure Lengend Snippet: Summary of the drug/compound libraries used in the antifungal drug repurposing (see also Table S1, Supplementary Materials ).

Article Snippet: 1547 or 1581 FDA-approved drug library , Johns Hopkins, USA Johns Hopkins Clinical Compound Library (JHCCL) version 1.0 , C. albicans, C. auris, C. krusei, C. parapsilosis, C. tropicalis , [ , ] .

Techniques: Drug discovery

Journal: Oncotarget

Article Title: Oncogenic KRAS sensitizes premalignant, but not malignant cells, to Noxa-dependent apoptosis through the activation of the MEK/ERK pathway

doi:

Figure Lengend Snippet: Best hits from the high-throughput screening. HeLa cells were treated with FDA-approved drugs in combination with SM83 and izTRAIL. The most effective 10 compounds enhancers of the cytotoxic effect are listed

Article Snippet: At day 2, media was changed with cells being exposed to 100 nM SM83 in addition to FDA-approved drug libraries (ENZO Life Sciences, MicroSource Discovery Systems Inc. and Prestwick Chemical, France) with the drug library compounds present at a final concentration of 1 μM.

Techniques:

Journal: Oncotarget

Article Title: Oncogenic KRAS sensitizes premalignant, but not malignant cells, to Noxa-dependent apoptosis through the activation of the MEK/ERK pathway

doi:

Figure Lengend Snippet: (A) The parental human epithelial (HME) cell line and the isogenic cell lines with knock-in mutations in KRAS (G13D), PI3K (H1047R) and EGFR (delE746A750) were treated with varying doses of CPT alone (left panel) or in combination with 100 nM SM83 (right panel). Viabilities are shown after 24 h of treatment. (B) HME D13/+ cells were pre-incubated with DMSO, 50 μM z-VAD, 20 μM Nec-1 (left panel), 10 μg/ml Infliximab (IFX, middle panel) and 10 μg/ml Enbrel (right panel) for 1 h and subsequently treated with 100 nM SM83 and 1 μM CPT. Cell viability was determined after 24 h. (C, D) HME +/+ and HME D13/+ cells were mock treated and treated with 100 nM SM83, 1 μM CPT and with their combination for 6 h. Cells were lysed and subjected to western blot to detect the apoptosis markers cleaved PARP, caspase-3 and caspase-8 (C) and the SM targets cIAP1, cIAP2 and XIAP (D). Actin is the loading control, asterisks show the cleaved forms p17/p19 of caspase-3 and the pro-caspase p55/p57 forms of caspase-8, together with its cleaved forms p41/p43. One representative of two independent experiments is shown.

Article Snippet: At day 2, media was changed with cells being exposed to 100 nM SM83 in addition to FDA-approved drug libraries (ENZO Life Sciences, MicroSource Discovery Systems Inc. and Prestwick Chemical, France) with the drug library compounds present at a final concentration of 1 μM.

Techniques: Knock-In, Incubation, Western Blot, Control

Journal: Oncotarget

Article Title: Oncogenic KRAS sensitizes premalignant, but not malignant cells, to Noxa-dependent apoptosis through the activation of the MEK/ERK pathway

doi:

Figure Lengend Snippet: (A) HME +/+ and HME D13/+ were transfected with siRNA targeting KRAS for 48 h and subsequently treated with 100 nM SM83 and 1 μM CPT. Cell viability was determined after 24 h of treatment. (B) HME pINDUCER20-Mock (Mock) and HME pINDUCER20-KRAS G13D (G13D) were incubated with doxycycline (Dox, 250 ng/ml) for 48 h, lysed and a western blot was performed. The presence of activated KRAS was determined by detection of phosphorylated ERK1/2. (C) Active GTP-RAS was purified in cells stimulated as in (B) by pull-down assay using the recombinant RBD domain of RAF1; HME D13/+ are shown as positive control for activated KRAS. (D) HME Mock and KRAS G13D were incubated with Dox (250 ng/ml) for 48 h and treated with 100 nM SM83 and 1 μM CPT. Cell viability was determined after 24 h. (E) MCF10A Mock and KRAS G13D were incubated with Dox (250 ng/ml) for the indicated time, lysed and analyzed by western blot for the detection of ERK1/2 and phosphorylated ERK1/2. Actin is shown as a loading control. (F) MCF10A Mock and KRAS G13D were incubated with Dox (250 ng/ml) for 48 h and treated with 100 nM SM83 and 0.1 μM CPT. Cell viability was determined after 24 h. One representative of two independent experiments is shown.

Article Snippet: At day 2, media was changed with cells being exposed to 100 nM SM83 in addition to FDA-approved drug libraries (ENZO Life Sciences, MicroSource Discovery Systems Inc. and Prestwick Chemical, France) with the drug library compounds present at a final concentration of 1 μM.

Techniques: Transfection, Incubation, Western Blot, Purification, Pull Down Assay, Recombinant, Positive Control, Control

Journal: Oncotarget

Article Title: Oncogenic KRAS sensitizes premalignant, but not malignant cells, to Noxa-dependent apoptosis through the activation of the MEK/ERK pathway

doi:

Figure Lengend Snippet: (A, left panel) HME +/+ and HME D13/+ cells were treated with 100 nM SM83 and 1 μM CPT for 6 h, lysed and subjected to western blot to detect Noxa and Mcl1 levels. (A, right panel) HME/MCF10A Mock and KRAS G13D were incubated with Dox (250 ng/ml) for the indicated time and subjected to western blot to detect Noxa and Mcl1 levels. Actin is shown as the loading control. (B) HME D13/+ cells were trasfected with control and Noxa-targeting siRNAs and, after 48 h, were treated with the indicated concentration of CPT (μM) for 6 h. Cells were then lysed and analyzed by western blot to evaluate Mcl1 levels. Noxa is shown to check the silencing efficiency and actin as the loading control. (C) HME +/+ and HME D13/+ were transiently transfected with siRNA targeting Noxa for 48 h and subsequently treated with 100 nM SM83 and 1 μM CPT. Cell viability was determined after 24 h. One representative of two independent experiments is shown.

Article Snippet: At day 2, media was changed with cells being exposed to 100 nM SM83 in addition to FDA-approved drug libraries (ENZO Life Sciences, MicroSource Discovery Systems Inc. and Prestwick Chemical, France) with the drug library compounds present at a final concentration of 1 μM.

Techniques: Western Blot, Incubation, Control, Concentration Assay, Transfection

Journal: Oncotarget

Article Title: Oncogenic KRAS sensitizes premalignant, but not malignant cells, to Noxa-dependent apoptosis through the activation of the MEK/ERK pathway

doi:

Figure Lengend Snippet: (A) HME +/+ and HME D13/+ cell lines were pre-incubated with 50 μM PD98059, 25 μM UO126, 20 μM Triciribine and 20 μM LY294002 for 2 h, and then treated with 100 nM SM83 and 1 μM CPT. Cell viability was quantified after 24 h. One representative of three independent experiments is shown. (B) HME +/+ (left panel) and HME D13/+ (right panel) cell lines were treated with 50 μM PD98059, 25 nM UO126, 20 μM Triciribine and 20 μM LY294002 for 2 h, and subsequently analyzed by western blot to detect the phosphorylated forms of AKT, ERK1 and ERK2, their total levels (upper panels) or Noxa and Mcl1 (lower panels). Actin is shown as loading control. (C) HME +/+ and HME D13/+ were transiently transfected with the indicated siRNAs for 72 hours and subsequently analyzed by western blot to detect total and phosphorylated ERK1 and ERK2, and their total levels (left panel), Noxa and Mcl1 (right panel). Actin is shown as loading control. (D) Parental HME +/+ and HME D13/+ cells were silenced for 48 h and then treated with DMSO and 100 nM SM83 plus 1 μM CPT for further 24 h. One representative of three independent experiments is shown. * P < 0.05 vs siCtr.

Article Snippet: At day 2, media was changed with cells being exposed to 100 nM SM83 in addition to FDA-approved drug libraries (ENZO Life Sciences, MicroSource Discovery Systems Inc. and Prestwick Chemical, France) with the drug library compounds present at a final concentration of 1 μM.

Techniques: Incubation, Western Blot, Control, Transfection

Journal: Oncotarget

Article Title: Oncogenic KRAS sensitizes premalignant, but not malignant cells, to Noxa-dependent apoptosis through the activation of the MEK/ERK pathway

doi:

Figure Lengend Snippet: SW48 +/+ and SW48 D13/+ (A), HCT116 +/− and HCT116 D13/− (B), Lim1215 +/+ and Lim1215 D13/+ (C), DLD1 D13/− and DLD1 +/− (D) cell lines were treated with DMSO and the combination of SM83 and CPT at varying concentrations. Cell viability was evaluated after 24 h. One representative of three independent experiments is shown. SW48 +/+ and SW48 D13/+ (E), HCT116 +/− and HCT116 D13/− (F), Lim1215 +/+ and Lim1215 D13/+ (G), DLD1 D13/− and DLD1 +/− (H) cell lines were treated with DMSO and 100 nM SM83, 0.1 μM CPT either alone or in combination for 6 h. Cells were lysed and analyzed by western blotting to determine Noxa and Mcl1 levels. One representative of two independent experiments is shown. Asterisk indicates the specific band of Actin shown as loading control.

Article Snippet: At day 2, media was changed with cells being exposed to 100 nM SM83 in addition to FDA-approved drug libraries (ENZO Life Sciences, MicroSource Discovery Systems Inc. and Prestwick Chemical, France) with the drug library compounds present at a final concentration of 1 μM.

Techniques: Western Blot, Control

Journal: Oncotarget

Article Title: Oncogenic KRAS sensitizes premalignant, but not malignant cells, to Noxa-dependent apoptosis through the activation of the MEK/ERK pathway

doi:

Figure Lengend Snippet: (A) HCT116 and (B) DLD1 cells were pre-incubated with 50 μM PD98059, 25 μM UO126, 20 μM Triciribine and 20 μM LY294002 for 2 h, and then mock-treated or treated with 100 nM SM83 and 1 μM CPT. Cell viability was quantified after 24 h. (C) HCT116 and (D) DLD1 cells were treated for 2 h with the indicated inhibitors as in (A) and then analyzed by western blot to detect Noxa levels and total and phosphorylated AKT and ERK levels. Actin is shown as loading control. (E, left panel) HCT116 D13/− cells were treated with 1 μM GDC-0941 for 2 h and analyzed by western blot to detect total and phosphorylated levels of AKT. Actin is shown as the loading control. (Right panel) Viability of HCT116 D13/− pre-treated with 1 μM GDC-0941 for 2 h and then treated with 100 nM SM83 and 1 μM CPT. Cell viability was quantified after 24 h and expressed as viability percentage to inhibitor alone. (F) HCT116 cells were transfected with siRNAs targeting ERK1 and ERK2, cells were collected after 72 h and analyzed by western blot to detect Noxa and total and phosphorylated levels of AKT and ERK. Actin is shown as loading control.

Article Snippet: At day 2, media was changed with cells being exposed to 100 nM SM83 in addition to FDA-approved drug libraries (ENZO Life Sciences, MicroSource Discovery Systems Inc. and Prestwick Chemical, France) with the drug library compounds present at a final concentration of 1 μM.

Techniques: Incubation, Western Blot, Control, Transfection