Labeling of CD38 in Raji cells. (A) Confocal
image of Raji cells
labeled with SR101–F-araNMN. Confocal microscope settings for
(A): laser power: 5.5%, pinhole: 1.1 airy units, master gain for PMT:
866. (B) Confocal image of Raji cells blocked with 6-alkyne-(F-araNAD)
then labeled with SR101–F-araNMN (visualization of intracellular
CD38 only). DAPI (blue) staining dsDNA showing nucleus. Confocal microscope
settings for (B): laser power: 10%, pinhole: 1.1 airy units, master
gain for PMT: 866. An increase in laser power and master gain was
necessary in order to have enough fluorescence emission to see the
signal. This indicates a low amount of intracellular active CD38.
(C) In-gel fluorescence analysis: lanes 1, live cell labeling with
Rh-6-(F-araNAD) followed by in-gel fluorescence (labeling plasma membrane
CD38 only); lanes 2, whole cell lysate was obtained first followed
by labeling with Rh-6-(F-araNAD) (labeling all catalytically active
CD38); lanes 3, whole cell lysate without CD38 probes. Ladder is on
the left, listed first. The full gel images are shown in Figure S7
in SI.