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Although both substance types inhibited NHE1, the toxic results had been NHE1-independent, as inhibitor-induced viability loss was unaffected by NHE1 CRISPR/Cas9 knockout. EIPA and HMA accumulated thoroughly in spheroids, and also this was related to marked vacuolization, obvious autophagic arrest, ER tension, mitochondrial- and DNA harm and poly-ADP-ribose-polymerase (PARP) cleavage, indicative of severe stress and paraptosis-like cell death. Pyrazinoylguanidine-induced cellular death ended up being