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. 2021 Sep 23;47(2):409–421. doi: 10.1007/s11064-021-03454-3

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© The Author(s) 2021

Open AccessThis 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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Fig. 3 — HIF-1α inhibition led to diminished levels of NEFL and a lower average neurite length. Cells were stained for immunofluorescence with anti-NEFL antibody (red) as a neuronal marker and anti-GFAP antibody (green) as a glial marker. A shows the undifferentiated (undiff.) cell morphology, B shows sSCR + RA cells, C correspond to sHIF + RA cells. D and E show the fluorescence intensity of NEFL and GFAP, respectively. sSCR + RA cells demonstrated greater levels of NEFL immunoreactivity in comparison to Undifferentiated and sHIF + RA cells. Undiff. cells displayed higher GFAP immunoreactivity than sSCR + RA and sHIF + RA. Each symbol represents the Corrected total cell fluorescence (CTCF) of a single cell. Error bars represent mean ± SD. F and G show the total amount of neurites/cells and the average neurite length, respectively. Undiff. cells demonstrated a lower amount of neurites than sSCR + RA. Furthermore, sSCR + RA cells displayed a higher neurite average length. Although, sHIF + RA cells had an average neurite length higher than undiff. cells. Columns represent mean ± SD (one-way ANOVA followed by Tukey’s Multiple Comparison post-hoc test *p < 0.05, **p < 0.005, ***p < 0.0003, ****p < o.0001)