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. 2020 May 15;9(5):51. doi: 10.1038/s41389-020-0231-2

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© The Author(s) 2020

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. 6 — a Transcriptional analysis of the nuclear encoded OXPHOS genes of PDAC cells treated with metformin and of the untreated control cells. Left side: bar plot of the fold change of genes between two groups. Red bars indicate increase and blue bars decrease in gene expression. Right side: heatmap of the expression z-scores of the genes among samples. b Effects of metformin on the expression of COX6B2 mRNA. qRT-PCR analysis provides the COX6B2 mRNA levels in PANC-1 cells after treating with metformin for 7 d. This experiment was performed in technical triplicates. c qRT-PCR analysis of COX6B2 mRNA levels in PANC-1 cells with or without metformin (10 mM). The time point for the culture of cells with α-amanitin (50 μM) were indicated. Before the addition of α-amanitin, cells were pretreated with or without metformin (10 mM) for additional 36 h. The experiment was performed in triplicates and was independently repeated (n = 2). d, e Western blot analysis of COX6B2 in PANC-1 (d) and PaTu-8988t (e) cells after metformin (10 mM) treatment for 1–6 d. f, g 2% DDM-permeabilized PANC-1 cells (f) and 8988 cells (g) analyzed by BNG show mitochondrial monomer complex II–V levels after metformin (10 mM) treatment for 2 days. Cells without metformin were used as control. Complex II–V were probed with anti-SDHA, anti-UQCRCR, anti-COI, and anti-ATP5A, respectively. h Wound healing assays of PaTu-8988t cells after treatment with various concentrations (0, 1, and 4 mM) of metformin. i Western blot analysis of COX6B2 (upper panel) and cell proliferation assays (lower panel) of PaTu-8988t cells after treatment with various concentrations (0, 0.5, 1, 2, 4, 6, and 8 mM) of metformin. All quantified data are presented as mean ± SEM (n ≥ 3). *P < 0.05, **P < 0.01, ***P < 0.001. NS no significance.