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

Journal: Cardiovascular Research

Article Title: E-cigarette exposure augments murine abdominal aortic aneurysm development: role of Chil1

doi: 10.1093/cvr/cvac173

Figure Lengend Snippet: Nicotine induces and augments pro-inflammatory cytokines and ROS in human AoSMC and murine RAW264.7 cells. (A) Nicotine increases expression levels of inflammatory genes (Ccl2, Il6 and Tnf) in murine RAW264.7 cells. (B) Nicotine dose-dependently augments inflammatory gene (Ccl2, Tnf) responses to recombinant IL-6 treatment (20 ng/mL) in RAW264.7 cells. (C) Nicotine dose-dependently increases total ROS in RAW264.7 cells (DCFDA Assay, Ex/Em = 485/535 nm), a process reversed by TEMPOL. (D) Nicotine dose-dependently increases expression levels of inflammatory genes (CCL2, IL6 and IL8) in human AoSMCs. (E) Nicotine dose-dependently augments inflammatory gene (CCL2, IL6 and IL8) responses to recombinant IL-6 treatment (20 ng/mL) in human AoSMCs. (F) Nicotine dose-dependently increases ROS response to IL6 (20 ng/mL) in human AoSMCs (Abcam/Rhodamine, Ex/Em = 550/620 nm). (G) Corresponding pictures to (F) of human AoSMCs stained for ROS under green channel using ROS Detection Assay (Abcam/Rhodamine). Gene expression data are qRT–PCR presented as fold change vs. control. §P < 0.05 vs. control; vs. §§ control and IL6; † vs. control, IL6, and IL6 + nicotine 10 nM (A, B, D, and E). *P < 0.05 vs. control; #P < 0.05 vs. other group (C). *P < 0.05 or **P < 0.01 vs. IL6, #P < 0.05 vs. other group (F). All two-tailed Student’s t-test. Data are mean ± SEM (n = 4–6/treatment group).

Article Snippet: The DCFDA Cellular Reactive Oxygen Species (ROS) Detection Assay Kit (Abcam/ab113851) was used to quantitatively measure ROS in adherent RAW264.7 cells after 24 h of treatment.

Techniques: Expressing, Recombinant, Staining, Detection Assay, Quantitative RT-PCR, Two Tailed Test