Journal: Hepatology Communications

Article Title: Genome-wide CRISPR screen identifies ACSL3 as a regulator of lipotoxicity and progression of MASLD

doi: 10.1097/HC9.0000000000000884

Figure Lengend Snippet: ACSL3 deficiency abrogates PA-induced activation of lipogenic programs without enhancing β-oxidation. (A–C) mRNA levels by RT-qPCR normalized to ACTB: CPT1, CPT2, and ACOX (A); SCD, ChREBP, and SREBP1 (B); PPARG and LXRA (C). Abbreviations: KO, knockout; WT, wild type.

Article Snippet: Primers used in the TaqMan assays (ThermoFisher Scientific) for ACSL3 (Hs00244853_m1), CPT1A ( Hs00912671_m1), CPT2 ( Hs00988962_m1), ACOX1 ( Hs01074241_m1), SCD ( Hs01682761_m1), ChREBP (MLXIPL; Hs00975714_m1), SREBP1 (SREBF1; Hs02561944_s1), PPARγ (Hs01115513_m1), and LXRα (Nr1h3; Hs00172885_m1 ) were also used.

Techniques: Activation Assay, Quantitative RT-PCR, Knock-Out

Journal: APL Bioengineering

Article Title: Integrating Mendelian randomization and multi-omics analysis unravels gut microbiota-driven metabolic mechanisms in sepsis and identifies diagnostic biomarkers through experimental validation

doi: 10.1063/5.0296018

Figure Lengend Snippet: Diagnostic value of key diagnostic genes. (a) ROC curves illustrating the diagnostic performance of the three key diagnostic genes. (b)–(d) Expression levels of ABCC1 , CYP1B1 , and PPARG in dataset GSE134364 . (e)–(g) Expression levels of ABCC1 , CYP1B1 , and PPARG in dataset GSE65682 . ROC: receiver operating characteristic curve; AUC: area under the ROC curve.

Article Snippet: The membrane was blocked and incubated overnight at 4 °C with primary antibodies: ABCC1 (A2223, Abclonal, 1:1000), CYP1B1 (A1377, Abclonal, 1:1000), and PPARG (16643-1-AP, Proteintech, 1:1000).

Techniques: Diagnostic Assay, Expressing

Journal: APL Bioengineering

Article Title: Integrating Mendelian randomization and multi-omics analysis unravels gut microbiota-driven metabolic mechanisms in sepsis and identifies diagnostic biomarkers through experimental validation

doi: 10.1063/5.0296018

Figure Lengend Snippet: Expression of key diagnostic genes in human alveolar organoids derived from pulmonary sepsis patients. (a) Representative bright-field image of human alveolar organoids cultured for 7 days; scale bar = 200 μ m. (b) HE staining of human alveolar organoids. (c) Immunofluorescence quantification of AQP5, SOX9, and SPC in human alveolar organoids. (d) WB analysis showing the expression levels of ABCC1, CYP1B1, and PPARG in human alveolar organoids.

Article Snippet: The membrane was blocked and incubated overnight at 4 °C with primary antibodies: ABCC1 (A2223, Abclonal, 1:1000), CYP1B1 (A1377, Abclonal, 1:1000), and PPARG (16643-1-AP, Proteintech, 1:1000).

Techniques: Expressing, Diagnostic Assay, Derivative Assay, Cell Culture, Staining, Immunofluorescence

Journal: APL Bioengineering

Article Title: Integrating Mendelian randomization and multi-omics analysis unravels gut microbiota-driven metabolic mechanisms in sepsis and identifies diagnostic biomarkers through experimental validation

doi: 10.1063/5.0296018

Figure Lengend Snippet: Expression of key diagnostic genes in bronchial organoids derived from pulmonary sepsis rats. (a) Representative bright-field image of rat bronchial organoids cultured for 7 days; scale bar = 200 μ m. (b) HE staining and IHC results for Ki67, P63, FOXJ1, MUC5AC, and CC10 in rat bronchial organoids. (c) Morphological status of rat bronchial organoids infected with four bacterial concentrations (MOI = 0, 5, 10, and 20) of Escherichia coli and Staphylococcus aureus at 0, 6, 12, and 24 h. (d) Fluorescence detection of organoid viability in rat bronchial organoids infected with MOI = 20 bacteria ( E. coli and S. aureus ) at 12 and 24 h. (e) RT-qPCR analysis quantifying the expression levels of ABCC1 , CYP1B1 , and PPARG in rat bronchial organoids.

Article Snippet: The membrane was blocked and incubated overnight at 4 °C with primary antibodies: ABCC1 (A2223, Abclonal, 1:1000), CYP1B1 (A1377, Abclonal, 1:1000), and PPARG (16643-1-AP, Proteintech, 1:1000).

Techniques: Expressing, Diagnostic Assay, Derivative Assay, Cell Culture, Staining, Infection, Fluorescence, Bacteria, Quantitative RT-PCR

Journal: APL Bioengineering

Article Title: Integrating Mendelian randomization and multi-omics analysis unravels gut microbiota-driven metabolic mechanisms in sepsis and identifies diagnostic biomarkers through experimental validation

doi: 10.1063/5.0296018

Figure Lengend Snippet: GSEA, immune infiltration, and GSVA analyses of key diagnostic genes. (a)–(c) GSEA analysis of ABCC1 , CYP1B1 , and PPARG , identifying enriched pathways associated with these genes. (d) Differences in immune infiltrating cell populations between healthy donors and sepsis patients. (e) Correlation analysis between ABCC1 , CYP1B1 , PPARG , and immune cells. (f) and (g) GSVA analysis of ABCC1 , CYP1B1 , and PPARG.

Article Snippet: The membrane was blocked and incubated overnight at 4 °C with primary antibodies: ABCC1 (A2223, Abclonal, 1:1000), CYP1B1 (A1377, Abclonal, 1:1000), and PPARG (16643-1-AP, Proteintech, 1:1000).

Techniques: Diagnostic Assay

Journal: APL Bioengineering

Article Title: Integrating Mendelian randomization and multi-omics analysis unravels gut microbiota-driven metabolic mechanisms in sepsis and identifies diagnostic biomarkers through experimental validation

doi: 10.1063/5.0296018

Figure Lengend Snippet: Single-cell analysis of ABCC1 , CYP1B1 , and PPARG . (a) and (b) Expression of ABCC1 , CYP1B1 , and PPARG in different immune cells.

Article Snippet: The membrane was blocked and incubated overnight at 4 °C with primary antibodies: ABCC1 (A2223, Abclonal, 1:1000), CYP1B1 (A1377, Abclonal, 1:1000), and PPARG (16643-1-AP, Proteintech, 1:1000).

Techniques: Single-cell Analysis, Expressing