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. 2021 Mar 4;11:5133. doi: 10.1038/s41598-021-84528-8

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Morphological analysis of mitochondria in Drp1 conditional knockout (cKO) and glucose deprivation (GD). (A) Representative images of primary Drp1^flx/flx neurons. Rows (top to bottom): control Drp1^flx/flx, Drp1^flx/flx after 18 hr GD, Drp1^flx/flx with Cre, Drp1^flx/flx with Cre after 18 hr GD. Untreated control groups are shown on rows 1 and 3, 18 h GD groups are shown on rows 2 and 4. Panels (from left to right): ATP synthase (green) and TOM20 (red) merged, MAP2 (magenta), DAPI (blue), all channels merged. Scale bars = 10 µm. (B) Percentage of mitochondrial area per morphology in Drp1 cKO and GD experiments. Mean ± SD (n = 5–8 per group). * indicates p < .05 versus Drp1^flx/flx. # indicates p < .05 versus Drp1^flx/flx + GD. $ indicates p < .05 versus Drp1^flx/flx + Cre. (C) Representative Western blot displaying Opa1 and GAPDH protein levels across Cre-Lentivirus concentrations. Full-length blots are presented in Supplementary Fig. 3. (D) Western blot quantification of Drp1 protein levels after Cre-Lentivirus treatment (n = 3–4). For all Drp1 cKO experiments, 2 × MOI Cre-Lentivirus was used.