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. 2019 Sep 5;76(22):4569–4580. doi: 10.1007/s00018-019-03290-3

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© The Author(s) 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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Fig. 1 — Differentiation induced expression of HPCA in SH-SY5Y cells. a, b SH-SY5Y cells were induced to differentiate by adding 50 μM all-trans retinoic acid (RA) for the indicated number of days. a The mRNA levels of the presynaptic marker, synaptophysin (SYP) and hippocalcin (HPCA), were determined by RT-qPCR. b Proteins were analyzed by western blotting with anti-HPCA, anti-SYP, and anti-calnexin antibodies. The graph shows mean densities as fold increases from five independent experiments (mean ± SEM). Band intensities were quantified using Quantity One^® software. **P < 0.01, ***P < 0.001 compared with day 0 of differentiation. c Neuronal differentiation was induced by adding 50 μM RA for the indicated number of days. Cells were stained with DAPI (blue) and an anti-SYP antibody (green) to visualize neurite extensions. Scale bar, 100 μm. d Neurite lengths were measured in randomly selected areas in three independent experiments. ***P < 0.001 compared with day 0 of differentiation (mean ± SEM)