Fig. 6: CA074 prevents eIF2α degradation and cell death
From: IRE1α deficiency promotes tumor cell death and eIF2α degradation through PERK dipendent autophagy
a,b U937 cells were treated with 4μ8 C (12.5 μM) + TN (3 μM) for 18 h and in the last 6 h also with or without 3-MA (5 mM), Bafilomycin (50 nM), Chloroquine (200 μM) or CA074 (10 μM). Cell death parameters were evaluated by calculating PI positive cells as percentage of total cells examined by cytofluorimetry a. A portion of these cells were fixed and stained with PI and subG1 events were evaluated in the cell cycle by cytofluorimetry b. For each parameter ≥ 10.000 events were acquired for each sample. The reported values are means ± S.D. (N = 3). Statistical analysis by Student’s t-test are shown. c Representative western blot for eIF2α detected in the lysates of U937 cells treated or not with 4μ8 C (12.5 μM) + TN (3 μM) for 18 h and with or without CA074 during the last 6 h. β-actin is shown as loading control. Specific primary antibodies and peroxidase-conjugated antibodies were used. The values under each band were obtained using the following formula: (densitometry value of the band under investigation / densitometry value of the band of the corresponding β-actin) / (densitometry value of the band under investigation in the lysate of untreated cells / the densitometry value of the β-actin band in the lysate of untreated cells). This experiment was repeated twice with comparable results
