Briefly, the mean arbitrary fluorescence intensity unit of each cell was first calculated based on the fluorescent intensity of the total pixels within the cell.
Fluorescence was measured on a BD FACS Canto II flow cytometer, and results are shown as geometric mean fluorescence expressed in arbitrary fluorescence units (AFU).
The threshold cycle (Ct) is the point where each kinetic curve reaches a common arbitrary fluorescence level (AFL), placed to intersect each curve in the region of exponential increase.
We also extend our methodology and demonstrate how to analyze trajectories that are not in protein numbers but in arbitrary fluorescence units, a more typical condition in experiments.
Fluorescence was expressed as arbitrary fluorescence units and was measured at the same instrument settings in all experiments.
Bands were identified according to their molecular weight and the arbitrary fluorescence intensity was calculated by Odyssey software.
Second, we do not have a calibration curve that allows us to calculate molecules of Gemini from arbitrary fluorescence data.
Caspase activity was estimated as the change in arbitrary fluorescence units normalized to milligrams of protein used in the assay.
The fluorescence intensity from samples was expressed as arbitrary fluorescence units, obtained at a wavelength of maximum emission.
Second, we do not have a standard calibration curve that allows us to calculate molecules of Gemini from arbitrary fluorescence data.
From this regression, we determined the enzymatic activity as the change in arbitrary fluorescence with respect to time (Figure S7 and Figure S8).
