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MCF-7 cells were cultured for 24 and 36 h without any compound (control) and with CA-4 or CA-4E at concentrations of 1.5 and 3 nM, respectively.
Apart from the basal biopsy, immediately frozen in liquid nitrogen, a second biopsy portion was cultured during 4 h without any compound (unchallenged), a third was incubated 4 h in the presence of 250 μg/ml PTG (PTG) and a last portion was challenged with the same amount of PTG after a 20-min incubation with 60 µg/ml Z (PTG-Z).
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If intervals without any compounds exist, the number of intervals is decreased by one and the binning method is reapplied.
To remove the 2-D bias, the five sets in Series B [designated as PubChem-(B), MeSH-(B), Protein3D-(B), PharmAct-(B), and Drug-(B)] were generated by using the unique set of parent compound representations in the respective sets in Series A and then excluding any compound without a computationally generated PubChem3D conformer description.
The Z score for metHb (Zmet) was also calculated according to eq 2. 2 In this equation, metHb%cpd is the metHb% of hemoglobin incubated with a unique compound, and metHb%ctrl is the metHb% of hemoglobin without any added compound.
Tubes without any test compound were used as a control.
The assay run without any test compound (containing only NBT riboflavine) was used as the reference.
The Z score for oxyHb% (Zoxy) was calculated for each compound according to eq 1. 1 In this equation, oxyHb%cpd is the oxyHb% of hemoglobin incubated with a unique compound, and oxyHb%ctrl is the oxyHb% of hemoglobin without any added compounds.
Cultures without any test compounds were taken as control.
Flexible self-standing films were obtained from the amphiphilic SQ-NPs without any additional compounds, which are due to intermolecular hydrogen bondings.
With the combustion/reduction furnaces and liquid nitrogen trap in the GC/C/IRMS system, the gas introduced to the IRMS ion source was much cleaner without any excess compounds but N2.
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