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. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: Cell Microbiol. 2014 May 1;16(9):1391–1404. doi: 10.1111/cmi.12299

Figure 2.

Figure 2

Immunofluorescence analysis of internalization of CdtB. THP-1 derived macrophages were exposed to media alone (black dotted line), CdtWT (red line) or CdtABCY71P (green line) for 1 hr and then analyzed by immunofluorescence and flow cytometry for the presence of CdtB following fixation, permeabilization and staining with anti-CdtB mAb conjugated to AlexaFluor 488. Panel A shows results of cells incubated at 37°C, fixed, permeabilized and stained for CdtB. Panel B shows results of cells incubated at 5°C and treated as described above. Panel C shows fluorescence associated with CdtB in non-permeabilized cells incubated at 37°C. Fluorescence is plotted versus relative cell number. Numbers represent the mean channel fluorescence (MCF); at least 10,000 cells were analyzed per sample. Results are representative of three experiments. The MCF for untreated cells (autofluorescent) was 4.