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The characterization of alcohol dehydrogenases in hALO-ALD models (A) Relative mRNA levels of ADH1B and ALDH1B1 in the control-treated and ethanol-treated hALOs were determined. The relative expression of each gene was normalized against that of 18S rRNA. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05 and ∗∗ p < 0.01. (B) The expression of ADH1B and ALDH1B1 was evaluated in the control-treated and ethanol-treated hALOs. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. (C) The expression of ADH1B and ALDH1B1 was evaluated in liver tissues from control individuals and three different patients with ALD. The histogram shows the difference in the percentages of ADH1B and ALDH1B1 in each field between control individuals and patients with ALD. The hash (#) represents a donor. There were 10 fields in each group. Scale bar, 50 μm. The results are presented as the mean ± SEM. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05 and ∗∗ p < 0.01. (D) The levels of cellular ROS in the control-treated and ethanol-treated hALOs were determined by a <t>DCFDA/H2DCFDA-Cellular</t> Reactive Oxygen Species Detection Assay Kit. The relative fluorescence intensity of ROS was determined using ImageJ software. Significant differences were determined by unpaired two-tailed paired t tests. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. (E) Cell viability in the control-treated and ethanol-treated hALOs was determined. The fluorescence intensity was determined using ImageJ software. Significant differences were determined by unpaired two-tailed paired t tests. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. ADH1B, alcohol dehydrogenase 1B; ALDH1B1, aldehyde dehydrogenase 1 family member B1.
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The characterization of alcohol dehydrogenases in hALO-ALD models (A) Relative mRNA levels of ADH1B and ALDH1B1 in the control-treated and ethanol-treated hALOs were determined. The relative expression of each gene was normalized against that of 18S rRNA. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05 and ∗∗ p < 0.01. (B) The expression of ADH1B and ALDH1B1 was evaluated in the control-treated and ethanol-treated hALOs. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. (C) The expression of ADH1B and ALDH1B1 was evaluated in liver tissues from control individuals and three different patients with ALD. The histogram shows the difference in the percentages of ADH1B and ALDH1B1 in each field between control individuals and patients with ALD. The hash (#) represents a donor. There were 10 fields in each group. Scale bar, 50 μm. The results are presented as the mean ± SEM. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05 and ∗∗ p < 0.01. (D) The levels of cellular ROS in the control-treated and ethanol-treated hALOs were determined by a DCFDA/H2DCFDA-Cellular Reactive Oxygen Species Detection Assay Kit. The relative fluorescence intensity of ROS was determined using ImageJ software. Significant differences were determined by unpaired two-tailed paired t tests. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. (E) Cell viability in the control-treated and ethanol-treated hALOs was determined. The fluorescence intensity was determined using ImageJ software. Significant differences were determined by unpaired two-tailed paired t tests. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. ADH1B, alcohol dehydrogenase 1B; ALDH1B1, aldehyde dehydrogenase 1 family member B1.

Journal: Cell Reports Methods

Article Title: Modeling alcohol-associated liver disease in humans using adipose stromal or stem cell-derived organoids

doi: 10.1016/j.crmeth.2024.100778

Figure Lengend Snippet: The characterization of alcohol dehydrogenases in hALO-ALD models (A) Relative mRNA levels of ADH1B and ALDH1B1 in the control-treated and ethanol-treated hALOs were determined. The relative expression of each gene was normalized against that of 18S rRNA. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05 and ∗∗ p < 0.01. (B) The expression of ADH1B and ALDH1B1 was evaluated in the control-treated and ethanol-treated hALOs. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. (C) The expression of ADH1B and ALDH1B1 was evaluated in liver tissues from control individuals and three different patients with ALD. The histogram shows the difference in the percentages of ADH1B and ALDH1B1 in each field between control individuals and patients with ALD. The hash (#) represents a donor. There were 10 fields in each group. Scale bar, 50 μm. The results are presented as the mean ± SEM. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05 and ∗∗ p < 0.01. (D) The levels of cellular ROS in the control-treated and ethanol-treated hALOs were determined by a DCFDA/H2DCFDA-Cellular Reactive Oxygen Species Detection Assay Kit. The relative fluorescence intensity of ROS was determined using ImageJ software. Significant differences were determined by unpaired two-tailed paired t tests. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. (E) Cell viability in the control-treated and ethanol-treated hALOs was determined. The fluorescence intensity was determined using ImageJ software. Significant differences were determined by unpaired two-tailed paired t tests. Scale bar, 50 μm. The results are presented as the mean ± SEM. n = 3. The asterisk (∗) represents statistical significance compared with the control: ∗ p < 0.05. ADH1B, alcohol dehydrogenase 1B; ALDH1B1, aldehyde dehydrogenase 1 family member B1.

Article Snippet: DCFDA/H2DCFDA-Cellular Reactive Oxygen Species Detection Assay Kit , Abcam , Cat# ab113851.

Techniques: Expressing, Detection Assay, Fluorescence, Software, Two Tailed Test

Journal: Cell Reports Methods

Article Title: Modeling alcohol-associated liver disease in humans using adipose stromal or stem cell-derived organoids

doi: 10.1016/j.crmeth.2024.100778

Figure Lengend Snippet:

Article Snippet: DCFDA/H2DCFDA-Cellular Reactive Oxygen Species Detection Assay Kit , Abcam , Cat# ab113851.

Techniques: Virus, Recombinant, Staining, Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Detection Assay, Quantitative RT-PCR, Software