Figure 7. Mitotic cells are phagocytosed... | Rockefeller University Press

Figure 7.

$Mitotic cells are phagocytosed more than their interphase neighbors. (A) Stills from Video 6 show an Her2 CAR GFP (green) macrophage phagocytosing a mitotic SKOV3 (mCherry-CAAX, magenta). (B) The graph shows the fraction of SKOV3 undergoing division in the entire population (left) compared with the fraction of phagocytosed SKOV3 cells undergoing division (right). (C) SKOV3 cells expressing FUCCI were treated with 0.1 μM paclitaxel or 1 μM STLC for 20 h to arrest cells in mitosis. Image shows cells in G1 phase (blue; hCdt1) and S/G2/M (magenta; hGeminin). (D) SKOV3 cells were treated with paclitaxel or STLC for 20 h, then the inhibitors were removed and replaced with fresh media and Her2 CAR GFP macrophages. The fraction of mCherry+ FUCCI SKOV3 cells (indicating S/G2/M phase) was measured for the entire SKOV3 population (total) and for the SKOV3 cells that were phagocytosed (of phagocytosed cells) during a 4-h time-lapse. (E) The graph shows the percent of Her2 CAR GFP or GFP-CAAX macrophages phagocytosing a cancer cell in the same experiment. (F) Raji B cells were treated with 0.1 μM paclitaxel or 1 μM STLC for 20 h to arrest cells in mitosis. The graph shows the percent of GFP-CAAX macrophages phagocytosing Raji cells opsonized with anti-CD47 antibody or with no antibody. For B, data were compared using a two sided Student’s T test and N = 3 independent experiments, consisting of ≥100 macrophages per replicate. For D–F, data were compared using one-way ANOVA with Holm–Sidak multiple comparison correction. N = 4 independent experiments, consisting of ≥100 macrophages per replicate. Bars represent the mean ± SEM. Data collected on the same day are annotated with the same shape point. * denotes P < 0.05; ** denotes P < 0.005; **** denotes P < 0.00005. Scale bars denote 20 μm. Refer to the image caption for details.$

Mitotic cells are phagocytosed more than their interphase neighbors. (A) Stills from Video 6 show an Her2 CAR GFP (green) macrophage phagocytosing a mitotic SKOV3 (mCherry-CAAX, magenta). (B) The graph shows the fraction of SKOV3 undergoing division in the entire population (left) compared with the fraction of phagocytosed SKOV3 cells undergoing division (right). (C) SKOV3 cells expressing FUCCI were treated with 0.1 μM paclitaxel or 1 μM STLC for 20 h to arrest cells in mitosis. Image shows cells in G1 phase (blue; hCdt1) and S/G2/M (magenta; hGeminin). (D) SKOV3 cells were treated with paclitaxel or STLC for 20 h, then the inhibitors were removed and replaced with fresh media and Her2 CAR GFP macrophages. The fraction of mCherry⁺ FUCCI SKOV3 cells (indicating S/G2/M phase) was measured for the entire SKOV3 population (total) and for the SKOV3 cells that were phagocytosed (of phagocytosed cells) during a 4-h time-lapse. (E) The graph shows the percent of Her2 CAR GFP or GFP-CAAX macrophages phagocytosing a cancer cell in the same experiment. (F) Raji B cells were treated with 0.1 μM paclitaxel or 1 μM STLC for 20 h to arrest cells in mitosis. The graph shows the percent of GFP-CAAX macrophages phagocytosing Raji cells opsonized with anti-CD47 antibody or with no antibody. For B, data were compared using a two sided Student’s T test and N = 3 independent experiments, consisting of ≥100 macrophages per replicate. For D–F, data were compared using one-way ANOVA with Holm–Sidak multiple comparison correction. N = 4 independent experiments, consisting of ≥100 macrophages per replicate. 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; **** denotes P < 0.00005. Scale bars denote 20 μm.