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. 2022 Aug 8;16:902278. doi: 10.3389/fncel.2022.902278

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Copyright © 2022 Duncan, Riordan, Hall, Payne, Chapman and Koulen.

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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Expression of N-acyl phosphatidylethanolamine phospholipase D (NAPE-PLD) in ReN cells under conditions of oxidative stress. (A) ReN cells express NAPE-PLD in the soma and nucleus. (B) Exposure of ReN cells to tert-butylhydroperoxide (tBHP) resulted in a biphasic NAPE-PLD expression pattern–2 μM tBHP led to a 20% increase in NAPE-PLD expression. Green fluorescence is NAPE-PLD and blue fluorescence is DAPI. (C,D) Western blot analysis reveals that the expression of a 48 kDa band (expected molecular weight is 45.6 kDa) corresponding to NAPE-PLD is upregulated 2.4-fold by exposure to 10 μM tBHP. (E,F) Colocalization analysis on image stacks reveals that there is little colocalization between NAPE-PLD and DAPI (Pearson’s R value < 0.20) except for when cells are exposed to 10 μM tBHP, a concentration that increases colocalization to Pearson’s R of 0.20. *p ≤ 0.05.