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. 2021 Jul 26;46:102082. doi: 10.1016/j.redox.2021.102082

Fig. 7.

Fig. 7

Glycolysis enhances cell retention of IR-780 via SLCO4A1. A-B Cellular IR-780 uptake was detected by flow cytometry (left). Quantification of average fluorescence intensity (middle). Representative images showing accumulation of IR-780 in lung fibroblasts (right). Cells were incubated with 0.5 μM IR-780 for 10 min at 37 °C in the dark, after removing free dye, cells were harvested for flow cytometry to detect fluorescence intensity. C-D Testing the effects of PPARA agonist (fenofibrate), glycolytic inhibitors (BAY876, 2-DG, 3PO) and OATPs inhibitor (BSP) on the uptake of IR-780 in myofibroblast (TGF-β1 induced). Cells were incubated with 0.5 μM IR-780 for 10 min at 37 °C in the dark, after removing free dye, the fluorescence of IR-780 was observed and images were captured by using a fluorescent microscope. E Volcano plots showing the results of genomic profiling of RNA from the GSE44723 dataset. The red or blue dots represent genes that are significantly (P < 0.05) up-regulated or down-regulated, respectively (n = 4 in each group); F Venn diagram showing that solute carrier family (SLC) genes and upregulated genes in GSE44723 shared 6 significantly upregulated SLC genes in rapid progressing pulmonary fibrosis compared with non-fibrotic individuals (P < 0.05 by t-test; fold change>2) (left). Heat map of genomic profiling of RNA showing the up-regulated SLCO4A1 (P < 0.05 by t-test; fold change>2) (right). G Relative mRNA (n = 3) and protein levels of SLCO4A1 in HFL1 treated with or without TGF-β1 and α-KG. H Representative immunoblots of the expressions of SLCO4A1 in control HFL1 or cells knockdown of SLCO4A1 (left). Representative images showing accumulation of IR-780 in HFL1 of indicated group (right). Drug doses: TGF-β1 (5 ng/ml), IR-780 (0.5 μM), BAY-876 (2.5 μM), 3-PO (5 mM), 2-DG (5 mM), fenofibrate (1 μM), BSP (250 μM), α-KG (1 mM). Bars represent 250 μm (A-D and H). The results are presented as the means ± S.D. (**P < 0.01; A and B, student's t-test; G, One-way ANOVA). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)