Toyocamycin

The IRE1α-XBP1 pathway inhibitors used were toyocamycin (Tocris Bioscience, Bristol, UK), 2-hydroxy-1-naphthaldehyde (HNA; Sigma-Aldrich), STF-083010 (Sigma-Aldrich), and 3-ethoxy-5 6-dilbromosalicylaldehyde (3ETH; Sigma-Aldrich). ES cells were seeded into 96-well plates at 5000 to 10000 cells/well. The next day, different concentrations of inhibitors or DMSO (as a vehicle control) were added to each well. After 72 h, the inhibitory effect of these inhibitors on the growth of ES cell lines was assessed using a Cell Counting Kit-8 (Dojindo JAPAN, Tokyo Japan) and a Microplate Reader (SAFIRE, TECAN, Männedorf, Switzerland). The IC50 was calculated using the GraphPad Prism program (GraphPad Software, Inc., CA USA).

Toyocamycin (Tocris Bioscience, Bristol, UK) was used as an IRE1α-XBP1 pathway inhibitor. GSK2606414 (S7307, Selleck) and ISRIB (trans-isomer; S7400, Selleck) were used as PERK pathway inhibitors. OS cells were seeded into 96-well plates at a density of 3000–10000 cells/well. The next day, different concentrations of inhibitors or DMSO (as a vehicle control) were added to each well. After 72 h, the inhibitory effect of these inhibitors on the growth of OS cell lines was assessed using a Cell Counting Kit-8 (Dojindo Japan, Tokyo, Japan) and a microplate reader (SAFIRE, TECAN, Männedorf, Switzerland). The IC50 was calculated using GraphPad Prism software version 9.2.0 (GraphPad Software, Inc., CA, USA).