G-Quadruplexes(GQs),which are formed by G-rich DNA sequences in human telomeres,have become an attractive target for cancer treatment.The ligands to stabilize the con-formation of human telomeric GQs in vivo are particularly important for structure-based ligand design and drug development targeting the noncanonical DNA structure.Here we report the conformational conversion of Tel26 induced by a naphthalene diimide(NDI)ligand in K+buffer,even at cellular physiological temperature(37℃)and under mimetic cellular crowding conditions created by Ficoll 70.We provide an insight into the dynamic conversion from initial hybrid-2 GQ topology to final parallel GQ topology.These results are helpful for the design of ligands with GQ conformation regulation.