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MWCNTs in media were diluted by culture media to final concentrations of 200, 50, and 12.5 μg/mL and added to cell cultures (in triplicate), respectively.
To investigate the impact of additional calcium within the exposure medium, calcium chloride was added to Jurkat media to final calcium concentrations of 2.1 mM or 5.1 mM.
The ATP standard was prepared with culture media to final volume of 40 uL, and 20 uL of ATPlite 1step reagent was added.
Doxorubicin (DOX) (AppliChem) was dissolved in water at stock concentration of 1 mg/ml and added to the culture media to final concentration of 2.5 μg/ml (4 μM) which corresponds to a therapeutic dosage [ 30].
HIV Nef protein (NIH AIDS Research and Reference Reagent Program) solutions were prepared in the same manner and diluted in media to final concentrations of 25, 50, 75 and 100 ng/ml.
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Proteins (at the indicated final concentrations) were diluted in media to a final volume of 50 µl and 2 µg N-[1- 2,3-Dioleoyloxy propyl]-N,N,N-trimethyl-ammonium methylsulfate (DOTAP) liposomal traN-[1- 2,3-Dioleoyloxy propyl]-N,N,N-trimethyl-ammoniumes.
Chloramphenicol was added to the media to a final concentration of 30.6 µg/mL or 106µg/mL.
Galactose was added into the media to a final concentration of 2% for continuing growth.
For cell proliferation assays, Alamar Blue dye (Invitrogen) was added directly into culture media to a final concentration of 10%.
Prior to MALDI-MS measurement, 18O NR was added to media to a final concentration of 10 µM as an internal standard.
Cells were collected at different time points after IL-22 was added to the culture media to a final concentration of 100 ng/ml.
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