Journal: JHEP Reports

Article Title: Increased serum miR-193a-5p during non-alcoholic fatty liver disease progression: Diagnostic and mechanistic relevance

doi: 10.1016/j.jhepr.2021.100409

Figure Lengend Snippet: Differences in miRNA levels detected in the NAFLD-only sequencing analysis .

Article Snippet: The expressions of 3 miRNAs, including the phosphorylated spike-in, were quantified using pre-formulated TaqManTM Advanced miRNA Assays (Thermo Fisher Scientific, Paisley, UK): 478293_mir (for the spiked-in cel-miR-39-3p), 477954_mir (hsa-miR-193a-5p), and 477855_mir (hsa-miR-122-5p).

Techniques: Sequencing, Activity Assay

Journal: JHEP Reports

Article Title: Increased serum miR-193a-5p during non-alcoholic fatty liver disease progression: Diagnostic and mechanistic relevance

doi: 10.1016/j.jhepr.2021.100409

Figure Lengend Snippet: miRNAs showing significantly different levels in NASH grouped by fibrosis stage .

Article Snippet: The expressions of 3 miRNAs, including the phosphorylated spike-in, were quantified using pre-formulated TaqManTM Advanced miRNA Assays (Thermo Fisher Scientific, Paisley, UK): 478293_mir (for the spiked-in cel-miR-39-3p), 477954_mir (hsa-miR-193a-5p), and 477855_mir (hsa-miR-122-5p).

Techniques:

Journal: JHEP Reports

Article Title: Increased serum miR-193a-5p during non-alcoholic fatty liver disease progression: Diagnostic and mechanistic relevance

doi: 10.1016/j.jhepr.2021.100409

Figure Lengend Snippet: Replication by qPCR of miR-193a-5p and miR-3687 associations. Levels of miR-193a-5p are shown for (A) significant fibrosis (NASH F2–F4) relative to minimal fibrosis (NAFL–NASH F0/F1) (n = 359), (B) advanced NAS (NAS 5–8) relative to mild NAS (NAS 1–4) (n = 359), and (C) advanced SAF activity (SAF activity 3-4) relative to mild SAF activity (SAF activity 0-2) (n = 359). Levels of miR-3687 are shown for (D) significant fibrosis (NASH F2–F4) relative to minimal fibrosis (NAFL–NASH F0/F1) (n = 371), (E) advanced NAS (NAS 5–8) relative to mild NAS (NAS 1–4) (n = 371), and (F) advanced SAF activity (SAF activity 3–4) relative to mild SAF activity (SAF activity 0–2) (n = 371). 2 -ΔΔCt was calculated relative to controls for all samples. Median values are shown with 95% CIs. Mann–Whitney U tests were performed for all comparisons. NAFL, non-alcoholic fatty liver; NAFLD, non-alcoholic fatty liver disease; NAS, NAFLD activity score; NASH, non-alcoholic steatohepatitis; qPCR, quantitative PCR; SAF, steatosis–activity–fibrosis.

Article Snippet: The expressions of 3 miRNAs, including the phosphorylated spike-in, were quantified using pre-formulated TaqManTM Advanced miRNA Assays (Thermo Fisher Scientific, Paisley, UK): 478293_mir (for the spiked-in cel-miR-39-3p), 477954_mir (hsa-miR-193a-5p), and 477855_mir (hsa-miR-122-5p).

Techniques: Activity Assay, MANN-WHITNEY, Real-time Polymerase Chain Reaction

Journal: JHEP Reports

Article Title: Increased serum miR-193a-5p during non-alcoholic fatty liver disease progression: Diagnostic and mechanistic relevance

doi: 10.1016/j.jhepr.2021.100409

Figure Lengend Snippet: Differentially expressed predicted target genes of miR-193a-5p in liver RNA-seq. (A) RNA-seq data from liver tissue were analysed for 3 comparisons: advanced NAS (NAS 5–8) relative to mild NAS (NAS 1–4), advanced SAF activity (SAF activity 3–4) relative to mild SAF activity (SAF activity 0–2), and significant fibrosis (NASH F2–F4) relative to minimal fibrosis (NAFL–NASH F0/F1). The genes that overlapped with those predicted by in silico tools to be targets of miR-193a-5p are shown in the heatmap. Hierarchical clustering is based on levels of fold change in gene expression in the liver tissue. Statistical significance in the RNA-seq data was determined using a Benjamini–Hochberg adjusted p value ≤0.05: ∗ p ≤0.05, ∗∗ p ≤0.01, and ∗∗∗ p ≤0.001. (B and C) Linear models of 80 overlapping NAFLD samples between the miRNA-seq and RNA-seq datasets for miR-193a-5p and (B) GPX8 and (C) COL1A1 . AFL, non-alcoholic fatty liver; NAFLD, non-alcoholic fatty liver disease; NAS, NAFLD activity score; NASH, non-alcoholic steatohepatitis; SAF, steatosis–activity–fibrosis.

Article Snippet: The expressions of 3 miRNAs, including the phosphorylated spike-in, were quantified using pre-formulated TaqManTM Advanced miRNA Assays (Thermo Fisher Scientific, Paisley, UK): 478293_mir (for the spiked-in cel-miR-39-3p), 477954_mir (hsa-miR-193a-5p), and 477855_mir (hsa-miR-122-5p).

Techniques: RNA Sequencing, Activity Assay, In Silico, Gene Expression

Journal: JHEP Reports

Article Title: Increased serum miR-193a-5p during non-alcoholic fatty liver disease progression: Diagnostic and mechanistic relevance

doi: 10.1016/j.jhepr.2021.100409

Figure Lengend Snippet: In vitro functional assessment of miR-193a-5p expression in Hep G2 cells. Hep G2 cells were treated with fatty acids (oleic acid [500 μM], palmitic acid [250 μM], or a combination of oleic [500 μM] and palmitic acid [250 μM]) for 24 h. All treatments were performed in triplicate, and qPCRs of each were performed in triplicate. Data were normalised to the control condition (untreated) using the 2 -ΔΔCt method. An unpaired Student’s t -test was performed for all conditions relative to the control conditions (∗∗ p ≤0.01). Data are presented as the mean with error bars representing the SEM. qPCR, quantitative PCR.

Article Snippet: The expressions of 3 miRNAs, including the phosphorylated spike-in, were quantified using pre-formulated TaqManTM Advanced miRNA Assays (Thermo Fisher Scientific, Paisley, UK): 478293_mir (for the spiked-in cel-miR-39-3p), 477954_mir (hsa-miR-193a-5p), and 477855_mir (hsa-miR-122-5p).

Techniques: In Vitro, Functional Assay, Expressing, Control, Real-time Polymerase Chain Reaction