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. 2021 Jul 8;26(10):6083–6099. doi: 10.1038/s41380-021-01207-w

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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 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. 4 — a IF of Hsp60 (green) and DAPI in AAV6 hSNCA/AAV6 PIAS2-mCherry- or AAV6 hSNCA/mutPIAS2-mCherry or CTR (mCherry) injected unilaterally into mouse brains. Scale bar is 14 um. b Quantification of Hsp60. Bars show mean ± SEM of n = 3; ^€P < 0.05 by ANOVA and **P < 0.01 by post hoc correction for multiple tests. c, d IF of Hsp60 (green), phosphorylated (p)α-Syn (red), nissl (yellow), and DAPI (blue) in brain of mice injected with AAV6 PIAS2-mCherry or AAV6 mCherry control. Scale bar is 14 um (c). d Quantification of Hsp60 (left) and pα-syn (right) and correlation between Hsp60 and pα-syn (below). Bars show mean ± SEM; *P < 0.05 by unpaired Student’s t-test. n = 3/group. Pearson’s correlation/simple linear regression test, n = 9/group; mCherry controls are in turquoise and PIAS2-mCherry is in purple. e IHC and quantification of 8OHdG in brains of WT mice overexpressing PIAS2-mCherry or CTR mCherry. Data represent mean number of positive nuclei and are presented as % of control. Scale bar, 250 μm. *P < 0.05 by unpaired Student’s t-test; n = 6/group. f IF for Tom20 and 8OHdG in striatum of WT mice overexpressing PIAS2-mCherry or CTR mCherry. Arrows indicate colocalization regions of 8OHdG and CoxI. Scale bars equal 10 microns. g IF for oxidized DJ1 in WT mice brain overexpressing mCherry or PIAS2-mCherry. Nuclei stained with DAPI. Scale bar 14 um. h WB of oxDJ1 and total DJ1 for CTR and PIAS2 expression. Vinculin was used as a loading control. Data represent % of control ± SEM of n = 3; ***P < 0.001 by Student’s t-test. i Expression of AAV6 PIAS2-mCherry or CTR (mCherry/yellow) in primary cortical neurons (CN) and IF staining for oxidized DJ1 (green) and nuclei stained with DAPI (blue). Scale bar, 16 μm. j, k WB of j oxidized DJ1 and total DJ1 and k pα-syn and total α-syn .k in CNs overexpressing PIAS2-mCherry or mCherry CTR. Vinculin was used as a loading control. Data represent % of control ± SEM; *P < 0.05 by Student’s t-test. n = 3. l WB for optin. and PINK1. H3 is used as a loading control. Data represent % of control ± SEM, *P < 0.05 by Student’s t-test. n = 3. m Staining for optin. and Tom20. n Quantification of l. WB of, o pERK1/2 and total ERK1/2, and p pP53 and total P53 in WT mouse brains overexpressing PIAS2-mCherry or mCherry CTR. Vinculin is used as a loading control. Data represent % of control ± SEM, *P < 0.05 by Student’s t-test. n = 3–4.