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. 2013 Feb 11;8(2):e55904. doi: 10.1371/journal.pone.0055904

Figure 3. Effect of γ-secretase inhibition on cellular viability in anchorage-independent culture conditions in relation with Hath1 and MUC2 expression.

Figure 3

A and B- Filter-grown HT29-Cl.27H or Caco2 cells, treated or not (control) with DBZ for 21 days, were resuspended and plated on polyHEMA-coated wells (nonadherent conditions) for 72 hours culture in 10% FCS containing medium without DBZ. A- RT-PCR detection of Hath1 and MUC2 mRNA: mean expression relative to control DMSO after normalization to actin gene expression; Mean ± SEM of 3 experiments; *, P<0.05; **, p<0.01. B- Relative cell viability versus control determined by trypan blue dye exclusion cell count. C- Filter-grown HT29-Cl.27H or Caco2 cells, pre-treated or not (control) with DBZ for 21 days were detached and seeded at 10, 000 cells per 60 mm dish in 0.35% agarose. The cells were maintained in culture for additional 15 days without supplementation of DBZ. Clusters of minimum 10 cells were counted. Mean relative expression to the control DMSO; Mean ± SEM of 3 experiments (3–4 dishes per experiment); ***, p<0.001. D- RT-PCR detection of Hes1 in Caco2 cells, treated or not (control) with DBZ for 21 days on filters: mean expression relative to control DMSO after normalization to actin gene expression; Mean ± SEM of 3 experiments.