Figure 4. Trogocytosis is more common... | Rockefeller University Press

Figure 4.

$Trogocytosis is more common than phagocytosis in multiple adherent cancer cell lines. Cancer cells were opsonized with a mouse anti-human CD47 antibody, and the amount of phagocytosis and trogocytosis was measured by flow cytometry. (A) Images show the double labeled (mCh-CAAX, H2B-iRFP) cancer cells used to distinguish phagocytosis and trogocytosis. (B) The cancer cell lines used were stained with a CD47 antibody and analyzed by flow cytometry. CD47 antibody binding was compared with an isotype control (gray). (C) Representative flow plots show the assay for distinguishing phagocytosis and trogocytosis by flow cytometry. (D) Graph shows the percent of mCherry-positive macrophages, indicating that these macrophages internalized some cancer cell material. (E) The graph shows the fraction of mCherry-positive macrophages that were also positive for iRFP, indicating cancer cell phagocytosis (gray), or lacking iRFP, indicating trogocytosis (pink). For D and E, data were compared using one-way ANOVA with Holm–Sidak multiple comparison correction. N = 4 independent experiments, consisting of three averaged technical replicates. Bars represent the mean ± SEM. Data collected on the same day are annotated with the same shape point. * denotes P < 0.05; ** denotes P < 0.005. Scale bar denotes 100 μm. Refer to the image caption for details.$

Trogocytosis is more common than phagocytosis in multiple adherent cancer cell lines. Cancer cells were opsonized with a mouse anti-human CD47 antibody, and the amount of phagocytosis and trogocytosis was measured by flow cytometry. (A) Images show the double labeled (mCh-CAAX, H2B-iRFP) cancer cells used to distinguish phagocytosis and trogocytosis. (B) The cancer cell lines used were stained with a CD47 antibody and analyzed by flow cytometry. CD47 antibody binding was compared with an isotype control (gray). (C) Representative flow plots show the assay for distinguishing phagocytosis and trogocytosis by flow cytometry. (D) Graph shows the percent of mCherry-positive macrophages, indicating that these macrophages internalized some cancer cell material. (E) The graph shows the fraction of mCherry-positive macrophages that were also positive for iRFP, indicating cancer cell phagocytosis (gray), or lacking iRFP, indicating trogocytosis (pink). For D and E, data were compared using one-way ANOVA with Holm–Sidak multiple comparison correction. N = 4 independent experiments, consisting of three averaged technical replicates. Bars represent the mean ± SEM. Data collected on the same day are annotated with the same shape point. * denotes P < 0.05; ** denotes P < 0.005. Scale bar denotes 100 μm.