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. 2024 Jun 25;28(1):326–339. doi: 10.1080/19768354.2024.2361144

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© 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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graphic file with name TACS_A_2361144_F0004a_OC.jpg — Regulation of cytokines following treatment with piperlongumine. HaCaT cells were treated with 1 μM piperlongumine at the indicated oxygen concentration. After incubation for 48 h, the clear supernatant was analyzed to detect specific cytokines and chemokines using enzyme-linked immunosorbent assays. (A) Representative immunoblot images and quantified results from whole membranes. (B) Map of specific cytokines and chemokines corresponding to each spot. (C) Statistically significant levels of secreted proteins are depicted as a heatmap analysis. The arbitrary value of the protein quantitation was normalized to that of vehicle-treated supernatant in 21% oxygen. (D) The expression of selective secretory proteins was analyzed. Positive and negative blots supplied by the manufacturer were used as the internal normalization control. GraphPad Prism was used to prepare the graphs, and statistical significance was determined using one-way ANOVA. ns; non-significant, *; p < 0.05, **; p < 0.01, ***; p < 0.005

graphic file with name TACS_A_2361144_F0004b_OB.jpg — Regulation of cytokines following treatment with piperlongumine. HaCaT cells were treated with 1 μM piperlongumine at the indicated oxygen concentration. After incubation for 48 h, the clear supernatant was analyzed to detect specific cytokines and chemokines using enzyme-linked immunosorbent assays. (A) Representative immunoblot images and quantified results from whole membranes. (B) Map of specific cytokines and chemokines corresponding to each spot. (C) Statistically significant levels of secreted proteins are depicted as a heatmap analysis. The arbitrary value of the protein quantitation was normalized to that of vehicle-treated supernatant in 21% oxygen. (D) The expression of selective secretory proteins was analyzed. Positive and negative blots supplied by the manufacturer were used as the internal normalization control. GraphPad Prism was used to prepare the graphs, and statistical significance was determined using one-way ANOVA. ns; non-significant, *; p < 0.05, **; p < 0.01, ***; p < 0.005