Dissecting stress-activated protein kinase (SAPK)-signaling pathways using multiplex gene knockout HeLa cells.

ABSTRACT: The stress-activated protein kinase (SAPK) family consists of three c-Jun N-terminal kinase (JNK) and four p38 members. To explore the isotype-specific or overlapping roles of SAPK members, HeLa-derived multiplex SAPK-KO cells, such as JNK1/2/3-triple KO, p38α/β/γ/δ-quadruple KO, and JNK1/2/3/p38α/β/γ/δ-septuple KO cells, were generated using the CRISPR-Cas9 method. Also, "sole survivor" (ss)-hextuple KO cells, in which only one of seven SAPK genes remains intact, were generated. Western blot analyses using phospho-specific antibodies for SAPK substrates showed that serum- or anisomycin-induced phosphorylation of MAPKAPK2, MSK1, Mnk1, and CREB (cyclic AMP response element-binding protein)/ATF-1 largely depended on p38, whereas anisomycin-induced phosphorylation of c-Jun/JunD depended on JN

SUBMITTER: Ito C 

PROVIDER: S-EPMC12704300 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature