CARS senses cysteine deprivation to activate AMPK for cell survival

A

RCC4 cells were cultured in complete medium for 24 h and then replaced and cultured with the corresponding amino acid‐deficient medium for 24 h, followed by WB detection of p‐AMPK and total AMPK protein expression. Actin served as the loading control.

B

WB analysis of p‐AMPK and total AMPK in 293T cells treated with DMSO, 1 µM Pim1 or 100 nM Takinib for 8 h under cystine deprivation. Actin served as the loading control.

C, D

293T cells were transfected with HA‐AMPKα1 alone or with Flag‐LKB1 (C) or Flag‐TAK1 (D) alone for 48 h and then cultured with cystine‐deficient medium for 8 h. Cell lysates were immunoprecipitated with anti‐Flag antibody, and a WB analysis was performed.

Figure EV1. Cystine starvation activates AMPK through CaMKK2 instead of LKB1 or TAK1.