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. 2021 Mar 2;11:4952. doi: 10.1038/s41598-021-83938-y

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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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Curcumin modulates phosphorylation of ERK by involving PPAR-γ activation. Immunofluorescence and western blot analysis of the time-course of 1 µM curcumin-induced ERK 1/2 phosphorylation are shown in (A) and (C). Representative photomicrographs of p-ERK immunofluorescence are shown in panel A, at the indicated time points. The upper panel in C shows a representative western blot of p-ERK and total ERK. To confirm the ERK 1/2 activation by curcumin (curc), OPs were pretreated for 30 min with 10 μM PD98059 (PD; ERK1/2 inhibitor) and p-ERK was evaluated by western blot (E). To evaluate the involvement of PPAR-γ in curcumin-induced ERK1/2 activation, OPs were pretreated with 1 μM GW9662 (GW; PPAR-γ antagonist; B and D). p-ERK was evaluated by IF (MFI value is shown in the lower panel in B. Data are mean ± SEM of n. 300 cells for condition) and by western blot (panel D; Data are means ± SEM of three independent experiments). *p < 0.0001 versus CTR; **p < 0.05 vs curc. Scale bar 30 µm. Image Lab 4.0 software (Bio-Rad, http://www.bio-rad.com/en-us/sku/1709690-image-lab-software) and Leica Application Suite Software (260RI, https: //www.leica-microsystems.com/products/microscope-software/details/product/leica-las-x-ls/) were used respectively for Wb and IF image acquisitions.