Figure 2
IGPR-1 promotes HT29 and HCT116 cells survival in suspension. Ectopic expression of IGPR-1 in HT29 and HCT116 cells is shown (a, b). HT29 and HCT116 cells expressing empty vector (pMSCV) or IGPR-1 were seeded in non-adherent 24-well plates (5 × 104/well) in 10%FBS in quadruple wells/group. Cell viability was determined by MTT assay at day 0, 2 and 4 (a, b). HT29 cells expressing empty vector or IGPR-1 were seeded in non-adherent six-well plates and survival of cells was measured by flow cytometry analysis by staining of cells with 7AAD-Annexin V. Live cells were defined as calcein violet+, 7AAD−, Annexin V−, and apoptotic/dying cells were defined as Annexin V+ (c). HT29 cells expressing empty vector or IGPR-1 were seeded in non-adherent 24-well plates for 24 h. Cells were lysed and whole-cell lysates were blotted for phospho-p38, total p38, IGPR-1 or for PLCγ1 as a protein loading control (d).
