Fig. 6.
Atf5 deficiency enhances the recovery of global translation under stress conditions. (A) Min6 cells were nucleofected with siRNA targeting Atf5 (siAtf5) or with nontargeting control siRNA (siNT). At 96 h after nucleofection, cells were treated with puromycin (1 μg/mL) for 30 min and then were harvested for protein. (Left) Western blot analysis with anti-puromycin for the basal level of global translation. (Right) Measurement of the relative density of blots. n = 3. (B) Min6 cells were nucleofected with siAtf5 or with siNT. At 96 h after nucleofection, cells were treated with Tg (1 μM) for the denoted time (in hours) and with puromycin (1 μg/mL) for the last 30 min and were harvested for protein. (Left) Western blot analysis with anti-puromycin for global translation by puromycin incorporation. (Right) Quantitation of relative density. n = 3. (C, Left) Islets harvested from Atf5+/+ and Atf5LacZ/LacZ mice were cultured, treated with Tg (1 μM) for the denoted time, and harvested for protein and Western blot analysis. (Right) Quantitation of relative density. n = 3. Bars show the mean, and error bars indicate the SEM. P values were calculated with Student’s t test; *P ≤ 0.05; **P ≤ 0.01.