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. 2018 Aug 7;9(61):31797–31811. doi: 10.18632/oncotarget.25737

Figure 2. The effect of loss of CD38 on cell proliferation and cell death in late stage tumors (age matched).

Figure 2

Paraffin and frozen sections or protein samples were prepared from WT and Cd38‒/‒ mice injected with B16F10 cells 28 days post injection. (A-D) Effect on proliferation. Proliferation was assessed using mitotic cell count in H&E stained sections (A and B) and Ki-67 staining (C and D). (A) Representative images of H&E stained tumors grown in WT and Cd38‒/‒ mice (scale bar = 100 μm); (arrows indicate representative mitotic cells). (B) Quantitation of the number of mitotic cells. The data presented are expressed as the mean ± SEM (bars) of the number of mitotic cells in mm2 (n = 10 mice from each group). n.s = not significant, Student’s t test. (C) Representative images of Ki-67 staining of tumors grown in WT and Cd38‒/‒ mice (scale bar = 200 μm) (D) Quantitation of the number of Ki-67 positive cells. The data presented are expressed as the mean ± SEM (bars) of the percentage of Ki-67-positive cells from total cells per field (n = 6 and 5 for WT and Cd38‒/‒ mice respectively). n.s = not significant, Student’s t test. Amount of cleaved (active) caspase-3 (Casp-3) in protein extracts prepared from the tumors (E, F). (E) A representative immunoblot probed for cleaved caspase-3 and β-tubulin. Negative control (NC) and positive control (PC) proteins were prepared from MEFs untreated or treated with staurosporine respectively. (F) Quantification of cleaved caspase-3 levels. Cleaved caspase-3 level is expressed as signal-intensity values (normalized to β-Tubulin). Expression levels were significantly higher in Cd38‒/‒ mice compared with WT (*p = 0.01; Student’s t test, n = 14 for each group). (G) Representative images of active caspase-3 staining in tumors originated from WT and Cd38‒/‒ mice (scale bar = 100 µm). (H) Quantification of cleaved caspase-3 levels. The data presented are expressed as the mean ± SEM (bars) of the average number of caspase-3 positive cells per field (n = 9 mice from each group). (*p = 0.02; Student’s t test).