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. 2023 Nov 6;6:241. doi: 10.1038/s42004-023-01043-9

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© The Author(s) 2023

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a MTT results acquired after cells exposure to 200 µM 6-OHDA for 30 min, 1 h, 2 h, and 3 h in RA/FBS(-) medium. The data is presented as a ratio of mean absorbance at 570 nm in the treated group (n = 6) compared to the control RA/FBS(-) group (n = 6). b Morphological analysis conducted after cells were exposed to 200 µM 6-OHDA for 30 min in RA/FBS(-) medium. cTyrosine hydroxylase expression assayed by ELISA with no RA: non-differentiated SH-SY5Y cells, RA/FBS(+): RA-differentiated cells. RA/FBS(-):RA-differentiated cells with 24 h FBS deficient medium incubation. 6-OHDA: RA/FBS(-) with 200 µM 6-OHDA for 30 min. d Mitochondrial membrane potential assay conducted with RA/FBS(-) cells exposed to 200 µM 6-OHDA for 30 min. The results are presented as a ratio of the mean green fluorescence intensity (FL1) divided by mean red fluorescent intensity (FL2) in the treated group (n = 3) compared to RA-control group (n = 3). e Reactive oxygen species (ROS) assay conducted with different concentrations (100 µM, 200 µM, 300 µM) of 6-OHDA for 30 min. The data are presented as a ratio of the mean fluorescent (FL1) intensity in the treated group (n = 3) compared to the RA/FBS(-) control group (n = 3). f Reactive oxygen species assay conducted with 200 µM of 6-OHDA for different incubation times (30 min, 1 h and 2 h). g Quantitative ELISA assay used to determine CREB phosphorylation after 200 µM 6-OHDA incubation for 30 min. The data are presented as the folded change of CREB phosphorylated protein (ng) per total protein (ng) in the treated group (n = 3) compared to the RA/FBS(-)control group (n = 3). n indicates the number of replicates per group and the error bars represent the standard deviation. The Dunnett test was used for multiple comparisons, and the Student t test was employed for comparing two groups, both using the Prism software. *P ≤ 0.05, **P ≤ 0.01, and ***P ≤ 0.001.