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. 2024 Oct 19;22:509. doi: 10.1186/s12964-024-01883-5

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.

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Fig. 7 — Pr-MPO induces cell death through Zn²⁺-mediated toxicity. A Effect of 5µM Pr-MPO treatment on HeLa cells using a 0% FBS culture medium supplemented with 1 or 2µM sodium acetate (NaAc) or zinc acetate (ZnAc). Statistical analysis was performed using One-way ANOVA followed by a post-hoc Dunnett’s multiple comparison test using the condition Pr-MPO 5µM as a control reference (** P < 0.01, *** P < 0.005). B Rescue effect of the Zn²⁺ chelator TPEN (0.5 and 2.5µM) on HeLa cells treated with 5µM Pr-MPO. Statistical analysis was performed using One-way ANOVA followed by a post-hoc Dunnett’s multiple comparison test using the untreated cell condition as a control reference (**** P < 0.0001). C Colony forming assay of untreated HeLa cells or treated with 5µM Pr-MPO, 500µM ZnAc, 2.5µM TPEN, 2.5µM TPEN + 5µM Pr-MPO or 2.5µM TPEN + 5µM Pr-MPO + 5µM ZnAc. D Western blot analysis of HeLa cells treated for 2 h with DMF, 5µM Pr-MPO and 500µM ZnAc; or DMF, 5µM Pr-MPO and 5µM Pr-MPO + 5µM in presence of 5µM TPEN; or DMF and 5µM Pr-MPO in presence of 0.1µM Bafilomycin A1; or DMF and 5µM Pr-MPO in presence of 500µM EDHB (iron chelator); or 200µM DCA (positive control) and 25µM LY30 (negative control). The level of Ser293 phosphorylation of PDH and total PDH was assessed as well as the autophagy marker LC3. This figure shows the results of the same samples run through two different gels: one gel was used to probe actin and S293PDH while the second gel was used to probe actin, total PDH and LC3; the use of two gels was necessary due to the strong resilience of the S293PDH antibody to stripping buffer. E Measure of total glucose, free glucose, glycogen and lactate in untreated HeLa cells or treated with 5µM Pr-MPO, 500µM ZnAc, 2.5µM TPEN, 2.5µM TPEN + 5µM Pr-MPO or 2.5µM TPEN + 5µM Pr-MPO + 5µM ZnAc. Statistical analysis was performed using One-way ANOVA followed by a post-hoc Dunnett’s multiple comparison test using the untreated cell condition as a control reference (* P < 0.05, ** P < 0.01, *** P < 0.005, **** P < 0.0001). F Measure of intracellular pH in untreated HeLa cells or treated with 5µM Pr-MPO, 500µM ZnAc, 2.5µM TPEN, 2.5µM TPEN + 5µM Pr-MPO or 2.5µM TPEN + 5µM Pr-MPO + 5µM ZnAc. Statistical analysis was performed using One-way ANOVA followed by a post-hoc Dunnett’s multiple comparison test using the untreated cell condition as a control reference (* P < 0.05, ** P < 0.01, *** P < 0.005)