Figure 8.

Characterisation of SMAD4 knockdown or knockout tumour derived cell lines. (A) Western blot of SMAD4 protein levels across cell lines re-derived from xenografts. (B) Relative cell number determined by a cell viability assay (AlamarBlue) at 144 hours following plating 2x10³ of CP-B sh1 SMAD4 knockdown (Control) cells or CP-B Ex1 SMAD4 knockout (Control) cells versus cells re-established from corresponding xenografts CP-B S1-2 and CP-B E1-1, respectively. Cell viability assays were performed on 3 independent occasions each with 6 technical replicates. Data points represent means of technical replicates for each individual experiment, bars represent mean ± SEM for N=3 experiments. ∗P < .05, ∗∗∗P < .001: Two-tailed Student t test. (C) Western blot of CDC6 protein across CP-B SMAD4 wild-type (Parental), SMAD4 knockout grown under in vitro conditions only (Ex1) cells and cell lines re-derived from SMAD4 knockdown (S1-1 – S1-3) and knockout (E1-1 – E1-3) tumor xenografts.