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Dihexa induces spinogenesis in cultured hippocampal neurons Actin-enriched spines increased in number in response to both dihexa (Figure 38B&D) and Nle'-AnglV (Figure 38C&D) following 5 days of treatment at 10 "12 M concentration that started on the 7 th day in vitro (DIV7) the results revealed a near 3-fold increase in the number of spines stimulated by dihexa and greater than 2-fold increase for Nle f -AngIV

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After a 12-min incubation at 25 °C, the temperature was elevated to 37 °C and maintained for 30 min, to promote macrocyclization