Figure 2: The AGN extract activated the integrated stress response (ISR) in HeLa cells. The HeLa cells were treated with the indicated concentrations of the AGN extract for 16 h (a) or with 10 μg/ml of the AGN extract for the indicated time periods (b). Cell lysates were subjected to immunoblot analyses using the indicated antibodies. The indicated fold increase in CHOP expression is the ratio of the CHOP to GAPDH. (c, d) The mRNA levels of ATF4 and CHOP were determined by real-time quantitative PCR. Statistical significance of the difference as calculated by Student’s t-test is with p<0.001.