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RA413S and RA414 induce both ER and oxidative stress triggering apoptosis.

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posted on 10.09.2021, 17:36 by Ravi K. Anchoori, Logan George, Ssu-Hsueh Tseng, Brandon Lam, Srinidhi Polkampally, Anjali D. Amiano, Palmer Foran, Hannah Tsingine, Harideep Samanapally, Fernanda Carrizo Velasquez, Samarjit Das, Deyin Xing, Ahmad Bin Salam, Balasubramanyam Karanam, Chien-Fu Hung, Richard B. S. Roden

(A) ES2 cells treated with compounds (0.5 μM) for 12 h and CHOP-10 mRNA levels, an ER stress marker, was analyzed by qRT-PCR. (B) ES2 cells were treated with compounds for 12 h and the total GSH level was assayed. (C) ES2 cells treated for 12 h with compounds (or as a positive control, H2O2) at the indicated doses, and then ROS levels were measured by adding Amplex Red and HRP. (D) To examine the early onset of apoptosis, ES2 cells were treated with compounds at indicated doses for 12 h then re-suspended in 100 μL binding buffer with 5 μL of Annexin V-PE and 5 μL of 7-AAD. After a 15 min incubation at RT, the cells were analyzed by flow cytometry using a FACSCalibur and CellQuest software.