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. 2012 Dec 5;7(12):e51044. doi: 10.1371/journal.pone.0051044

Figure 5. Arresten increases apoptosis of HSC-3 carcinoma cells in the organotypic model.

Figure 5

A. Organotypic sections were stained for the proliferation marker Ki-67 (brown). B. Proliferation was defined as a percentage of Ki-67-positive cells among the total number of carcinoma cells per microscopic field (200×magnification; n = total number of fields analyzed, 3–5 fields per organotypic section). C–F. Apoptotic cells were detected by TUNEL assay (green) and caspase-3 staining (red). Apoptotic cell death was quantified in terms of TUNEL (D) and caspase-3-positive (F) cells as a percentage of total number of carcinoma cells per microscopic field (200×magnification; n = total number of fields analyzed, 3–5 fields per organotypic section). Mann-Whitney U-test, ***p<0.001, *p<0.05. G. 20 µg of total protein of lysed cell extracts was separated by SDS-PAGE and immunoblotted with antibodies against signaling molecules of the Bcl-family apoptosis pathway, anti-apoptotic Bcl-xL and pro-apoptotic Bax. β-actin was used as a loading control. H. The relative band intensities were quantified (n = 3 Western analyses from separate protein extractions; mean ± SEM). Students t-test, *p<0.05.