Fig. 6

Overexpression of eNOS in cardiomyocytes increased NO content and inhibited the proliferation of lower neuronal cells. (A) NO content in the supernatant of the coculture model after PA stimulation in the upper cardiomyocytes overexpressing eNOS. (B) WB detection of P21, PGP9.5, and TH protein expression in lower layer PC12 neurons after PA stimulation in upper layer cardiomyocytes overexpressing eNOS. (C) EdU assay of proliferation levels of lower layer PC12 neuronal cells after PA stimulation in an upper layer cardiomyocyte coculture model overexpressing eNOS. Scale bar = 100 μm. (D) NO content in the supernatant of the coculture model after PA stimulation in the upper layer of cardiomyocytes overexpressing eNOS following the addition of C-PTIO to remove NO. (E, F) Following the addition of PA to the coculture system, eNOS was observed to be overexpressed in the upper chamber of H9C2 cells. Afterwards, CPTIO was introduced to eliminate NO, and WB analysis was conducted to measure the protein expression levels of P21, PGP9.5, and TH in PC12 cells located in the lower chamber. PC12 cell proliferation was evaluated using the EdU assay (20X). Scale bar = 100 μm