Figure 4.

Attenuation of UPR by cycloheximide (CHX) or sodium 4-phenylbutyrate (4-PBA) protects transformed cells from death. Cell death and UPR activation were analyzed in normal and transformed cells grown in LG for 72 h and treated for the further 24 h with CHX or 4-PBA. Normal (a and c) and transformed (b and d) cells were counted at 72 h and 96 h, after treatment with CHX (a and b) or 4-PBA (c and d). Data represent the average of at least three independent experiments (±S.D.); **P<0.01, Student's t-test. Phase contrast microscopy images of untreated and treated transformed cells at 96 h of culture are shown. (e–h) FACS analysis of normal (e and f) and transformed (g and h) cells stained with Annexin V-FITC and propidium iodide. The percentage of cell death for normal and transformed cells was calculated considering Annexin V- and PI-stained cells alone and in combination; representative dot plots of normal (f) and transformed (h) cells are shown. Data represent the average of at least three independent experiments (±S.E.M); *P<0.05, ***P<0.001, Student's t-test. UPR activation after CHX (i) and 4-PBA (j) treatments was followed through the expression analysis of Grp78 and CHOP proteins. Figures are representative of three independent experiments