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. 2023 Nov 13;16:1241420. doi: 10.3389/fnmol.2023.1241420

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Copyright © 2023 Kropf, Shekari, Jaberi, Puri, Wu and Fahnestock.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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BFCNs aged in vitro exhibit increased nitric oxide levels compared to young BFCNs. Rat BFCNs were stained with 5 μM DAF-FM for 1 h prior to imaging the cell bodies. Aged BFCNS [(B), DIV18-22] exhibit increased DAF-FM fluorescence (green) relative to young BFCNs [(A), DIV5-8], indicating that nitric oxide levels increase with in vitro age. Nuclei are labeled with DAPI (blue). (C) Quantification of the images shown in A,B, Student’s t-test. n = 30 images/group from three chambers in three independent experiments. The highest point in the DIV18-22 group was identified as an outlier using ROUT (Q = 1%). Removal of the outlier did not change the significance of the comparison. Error bars represent SEM. ****p < 0.0001. DIV, days in vitro. Scale bars: 10 μm.