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. 2022 Oct 14;13:6080. doi: 10.1038/s41467-022-33749-0

Fig. 3. Production of kynurenine by alcohol and its effect on autophagy flux.

Fig. 3

a Bar graph based on KEGG analysis to identify functional processes controlled by alcohol treatment using the same data as in Fig. 1a. FDR and genes are presented as a bar graph (FDR <0.05). b–d qRT-PCR, immunoblots, and immunohistochemical analyses for IDO and TDO2 in the liver of mice fed with the control diet or a Lieber-DeCarli alcohol liquid diet for 4 weeks (n = 5 each). Representative images were shown for (d). Scale bar: 100 μm. e sPLS-DA score plots based on the metabolomics data related to amino acids using the liver (left) and serum (right) from the mice fed either a control diet or a Lieber-DeCarli diet for 5 weeks (n = 7, 9 mice). f Heatmap analyses of increased amino acids and their metabolites using the same data as in (e) (n = 7, 9 mice; FDR <0.05; darker blue, stronger downregulation; darker red, stronger upregulation). Kynurenine was marked with red asterisks. g Concentrations of kynurenine and tryptophan, and kynurenine/tryptophan ratios using the same data as in (e) (n = 7, 9 mice). h Immunoblots for LC3BI/II and p62 in mPHs (left) or AML-12 cells (right) treated with 100 μM kynurenine for 12 h and continuously with 10 μM CQ for 2 h (repeated three times with similar results). i Immunoblots for LC3BI/II and p62 in mPHs treated with 100 μM kynurenine for 12 h (left) or AML-12 cells treated with 100 μM kynurenine for 12 h after transfection with siAhR (or siCon) for 24 h (right) (repeated three times with similar results). j Representative confocal microscopic images of staining for mCherry/GFP-LC3B puncta (upper) and their quantifications (lower) (n = 3 each, 10 cells/group for each experiment). mPHs were infected with Ad-mCherry-GFP-LC3B for 12 h and then exposed to 100 μM kynurenine (or vehicle) for 12 h. Scale bar: 10 μm. Values are expressed as means ± SEM. Significantly different compared to Con, Veh, or WT. Data were analyzed via two-tailed Student’s t-test (b, c, g) or one-way ANOVA with LSD (j). Source data are provided as a Source Data file.