Fig. 2: IRE1α impairment affects eIF2α expression
From: IRE1α deficiency promotes tumor cell death and eIF2α degradation through PERK dipendent autophagy
a Representative western blots out of three experiments are shown for p-eIF2α, t-eIF2α, ATF4 and CHOP detected in the lysates of U937 cells treated for 18 h with none, TN (3 μM) or 4μ8 C (12.5 μM) + TN (3 μM) for the indicated hours. Each protein was probed with specific antibody followed by peroxidase-conjugated secondary antibodies. β-actin is shown as loading control. b, c, d, e Quantitative analysis of each protein was performed by densitometry of the relative bands in 3 experiments. Values were obtained using the following formula: (densitometry value of the band under examination / densitometry value of the band of the corresponding β-actin)/(densitometry value of the band under examination in the control / densitometry value of the β−actin band in the control). Values are means ± S.D. The statistical differences (Student’s t-test) between the values obtained in 4μ8 C + TN cell treatments v/s the values obtained in the corresponding TN cell treatments are indicated as: *P < 0.05, **P < 0.01, ***P > 0.05. f IRE1α impairment through specific silencing affects eIF2α expression. U937 treated for 72 h with equal amount of scrambled siRNA or IRE1α specific siRNA were treated or not with TN (3 μM) for 18 h. t-eIF2α and CHOP were analyzed by western blot in the cell lysates by specific antibodies followed by peroxidase-conjugated antibodies. β-actin is shown as loading control and also to confirm the specificity of the transfected siRNA. Values under each band were obtained using the following formula: (densitometry value of the band under examination / densitometry value of the band of the corresponding β-actin) / (densitometry value of the band of the protein under examination in the lysate of scrambled treated cells / densitometry value of the β-actin band in the lysate of scrambled treated cells)
