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. 2014 Mar 24;136(15):5656–5663. doi: 10.1021/ja411046j

Figure 6.

Figure 6

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.