anti idi1  (Proteintech)

Journal: bioRxiv

Article Title: Magnesium isoglycyrrhizinate alleviates alcohol-associated liver disease through targeting HSD11B1

doi: 10.1101/2025.10.02.679991

Figure Lengend Snippet: (A) Chemical structure of MgIG. (B) Flowchart illustrating the modeling process for the NIAAA model. (C) Representative results of H&E and Oil Red O staining from the livers of mice in Ctrl, A-Ctrl and MgIG groups (n = 5). (D) Alterations in NAS (NAFLD activity score) and Oil Red O quantification (a.v.: arbitrary value) (n = 5). (E) Ratios of liver weight to body weight (LW/BW) in mice (n = 5). (F) Alterations in serum biochemical parameters (ALT, AST and TG) in mice in three groups (n = 5). (G) Alterations in mRNA expression of lipid metabolism genes ( Srebp-1c , Srebp2 , Acc1 , and Scd1 ), systemic inflammation markers ( Tnf-α , Il-6, Il-6, and Il-16 ), and apoptosis-related genes ( Bax and Bcl2 ) in the mice liver. The data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Scale bar: 50 μm.

Article Snippet: Antibodies against IDI1 (#11166-2-AP) and SREBP2 (#28212-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Staining, Activity Assay, Expressing

Journal: bioRxiv

Article Title: Magnesium isoglycyrrhizinate alleviates alcohol-associated liver disease through targeting HSD11B1

doi: 10.1101/2025.10.02.679991

Figure Lengend Snippet: (A, B) Co-IP was used to verify the direct binding between HSD11B1 and SREBP2 with or without MgIG treatment. (C, D) Alterations in protein levels of HSD11B1, p-SREBP2, n-SREBP2, and IDI1 in AML-12 cells induced by EtOH/PA, following knockdown or overexpression of Hsd11b1 , with and without MgIG treatment (n = 3). (E) Immunofluorescent staining was conducted to visualize the distribution and expression of SREBP2 (red) in EtoH/PA-induced AML-12 cells, with or without MgIG, following Hsd11b1 knockdown or overexpression. (F) Co-IP was used to verify the direct binding between HSD11B1 and IDI1. (G) Effects of Srebp2 on Idi1 transcriptional regulation were measured by luciferase assays in AML-12 and 293T cell lines. Idi1-wild-type (WT) or Idi1-mutant (Mut), plasmids with WT promoter cDNA clone of Idi1 or with mutant promoter cDNA clone plasmid; pRL-TK, an internal control reporter plasmid. (H, I) The expression levels of Hsd11b1 , Srebp2 , and Idi1 in normal liver were measured following the knockdown of Hsd11b1 , Srebp2 , and Idi1 , or the overexpression of Hsd11b1 and Idi1 , respectively (n = 4). (J, K) The expression levels of Hsd11b1 , Srebp2 , and Idi1 in ALD liver were measured following the knockdown of Hsd11b1 , Srebp2 , and Idi1 , or the overexpression of Hsd11b1 and Idi1 , respectively (n = 5). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001. **** P < 0.0001. Scale bar: 10 μm.

Article Snippet: Antibodies against IDI1 (#11166-2-AP) and SREBP2 (#28212-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Co-Immunoprecipitation Assay, Binding Assay, Knockdown, Over Expression, Staining, Expressing, Luciferase, Mutagenesis, Plasmid Preparation, Control

Journal: bioRxiv

Article Title: Magnesium isoglycyrrhizinate alleviates alcohol-associated liver disease through targeting HSD11B1

doi: 10.1101/2025.10.02.679991

Figure Lengend Snippet: (A-D) Representative liver H&E and Oil Red O staining results from ALD mice with Hsd11b1 , Srebp2 , or Idi1 knockdown or overexpression, with and/or without MgIG co-treatment. Changes in quantitative NAS (NAFLD activity score) and Oil Red staining (area %) were calculated and analyzed (n = 5). (E-F) Changes of serum ALT and TNF-α from ALD mice with Hsd11b1 , Srebp2 , or Idi1 knockdown or overexpression, with and/or without MgIG co-treatment (n = 5). (G) Alterations in protein levels of HSD11B1, p-SREBP2, n-SREBP 2, and Idi1 in ALD mice, following knockdown or overexpression of Hsd11b1 , with and without MgIG co-treatment (n = 3). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Scale bar: 50 μm.

Article Snippet: Antibodies against IDI1 (#11166-2-AP) and SREBP2 (#28212-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Staining, Knockdown, Over Expression, Activity Assay

Journal: bioRxiv

Article Title: Magnesium isoglycyrrhizinate alleviates alcohol-associated liver disease through targeting HSD11B1

doi: 10.1101/2025.10.02.679991

Figure Lengend Snippet: (A) Quantitative PCR validation of RNA-seq results revealed changes in Idi1 gene expression in AML-12 cells treated with ethanol/PA, with or without 0.25 mg/ml MgIG co-treatment (n = 4). (B) Quantitative PCR results confirmed the knockdown efficiency of Idi1 genes by siRNA in AML-12 cells. The changes in AML-12 cell viability following Idi1 siRNA transfection (24 h) were also assessed (n = 4). (C) Quantitative PCR results confirmed the overexpression efficiency of Idi1 genes via plasmid transfection in AML-12 cells. Changes in AML-12 cell viability following Idi1 plasmid transfection (24 h) were also evaluated (n = 4). (D) Changes in cell viability and apoptosis ratios were assessed in ethanol/PA-treated AML-12 cells, with or without 0.25 mg/ml MgIG, following Idi1 knockdown/overexpression (n = 4). (E, F) Western blot results for TNF-α, IL-6, Bax, and Bcl-2 and cell supernatant results for TNF-α and IL-6 in AML-12 cells treated with ethanol/PA and ethanol/PA + MgIG, with or without Idi1 knockdown/overexpression (n = 3). (G, H) Nile Red staining area (%) with corresponding representative cell staining images. Data are expressed as mean ± SD. For Western blot quantification: * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bar: 20 μm.

Article Snippet: Antibodies against IDI1 (#11166-2-AP) and SREBP2 (#28212-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Real-time Polymerase Chain Reaction, Biomarker Discovery, RNA Sequencing, Gene Expression, Knockdown, Transfection, Over Expression, Plasmid Preparation, Western Blot, Staining

Journal: bioRxiv

Article Title: Magnesium isoglycyrrhizinate alleviates alcohol-associated liver disease through targeting HSD11B1

doi: 10.1101/2025.10.02.679991

Figure Lengend Snippet: (A) Quantitative PCR validation of RNA-seq results revealed changes in Hsd11b1 gene expression in AML-12 cells treated with ethanol/PA, with or without 0.25 mg/ml MgIG co-treatment (n = 4). (B, C) Quantitative PCR confirmed the knockdown efficiency of Hsd11b1 (siRNA) and the overexpression efficiency of Idi1 (plasmid) in AML-12 cells, and cell viability changes were assessed 48 h after transfection (n = 4). (D) Nile Red staining area (%) with corresponding representative cell staining images (n=4). (E, F) Western blot results for TNF-α, IL-6, Bax, and Bcl-2 and cell supernatant results for TNF-α and IL-6 in AML-12 cells treated with ethanol/PA and ethanol/PA + MgIG, with or without Hsd11b1 knockdown/overexpression (n = 3). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bar: 20 μm.

Article Snippet: Antibodies against IDI1 (#11166-2-AP) and SREBP2 (#28212-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Real-time Polymerase Chain Reaction, Biomarker Discovery, RNA Sequencing, Gene Expression, Knockdown, Over Expression, Plasmid Preparation, Transfection, Staining, Western Blot

Journal: bioRxiv

Article Title: Magnesium isoglycyrrhizinate alleviates alcohol-associated liver disease through targeting HSD11B1

doi: 10.1101/2025.10.02.679991

Figure Lengend Snippet: (A, B) Co-IP was used to verify the direct binding between HSD11B1 and SREBP2 with or without MgIG treatment. (C, D) Alterations in protein levels of HSD11B1, p-SREBP2, n-SREBP2, and IDI1 in AML-12 cells induced by EtOH/PA, following knockdown or overexpression of Hsd11b1 , with and without MgIG treatment (n = 3). (E) Immunofluorescent staining was conducted to visualize the distribution and expression of SREBP2 (red) in EtoH/PA-induced AML-12 cells, with or without MgIG, following Hsd11b1 knockdown or overexpression. (F) Co-IP was used to verify the direct binding between HSD11B1 and IDI1. (G) Effects of Srebp2 on Idi1 transcriptional regulation were measured by luciferase assays in AML-12 and 293T cell lines. Idi1-wild-type (WT) or Idi1-mutant (Mut), plasmids with WT promoter cDNA clone of Idi1 or with mutant promoter cDNA clone plasmid; pRL-TK, an internal control reporter plasmid. (H, I) The expression levels of Hsd11b1 , Srebp2 , and Idi1 in normal liver were measured following the knockdown of Hsd11b1 , Srebp2 , and Idi1 , or the overexpression of Hsd11b1 and Idi1 , respectively (n = 4). (J, K) The expression levels of Hsd11b1 , Srebp2 , and Idi1 in ALD liver were measured following the knockdown of Hsd11b1 , Srebp2 , and Idi1 , or the overexpression of Hsd11b1 and Idi1 , respectively (n = 5). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001. **** P < 0.0001. Scale bar: 10 μm.

Article Snippet: Antibodies against IDI1 (#11166-2-AP) and SREBP2 (#28212-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Co-Immunoprecipitation Assay, Binding Assay, Knockdown, Over Expression, Staining, Expressing, Luciferase, Mutagenesis, Plasmid Preparation, Control

Journal: bioRxiv

Article Title: Magnesium isoglycyrrhizinate alleviates alcohol-associated liver disease through targeting HSD11B1

doi: 10.1101/2025.10.02.679991

Figure Lengend Snippet: (A-D) Representative liver H&E and Oil Red O staining results from ALD mice with Hsd11b1 , Srebp2 , or Idi1 knockdown or overexpression, with and/or without MgIG co-treatment. Changes in quantitative NAS (NAFLD activity score) and Oil Red staining (area %) were calculated and analyzed (n = 5). (E-F) Changes of serum ALT and TNF-α from ALD mice with Hsd11b1 , Srebp2 , or Idi1 knockdown or overexpression, with and/or without MgIG co-treatment (n = 5). (G) Alterations in protein levels of HSD11B1, p-SREBP2, n-SREBP 2, and Idi1 in ALD mice, following knockdown or overexpression of Hsd11b1 , with and without MgIG co-treatment (n = 3). Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Scale bar: 50 μm.

Article Snippet: Antibodies against IDI1 (#11166-2-AP) and SREBP2 (#28212-1-AP) were obtained from Proteintech (Wuhan, China).

Techniques: Staining, Knockdown, Over Expression, Activity Assay

Journal: bioRxiv

Article Title: Abnormal shear stress induces ferroptosis in endothelial cells via KLF6 downregulation

doi: 10.1101/2025.09.10.675441

Figure Lengend Snippet: RNA-seq reveals LSS suppresses cholesterol biosynthesis and UPR. (A) Volcano plot of DEGs (LSS vs. MSS;|log2FC|>0.6, Q<0.05); (B) KEGG pathways; (C) GO biological processes; (D) GSEA of steroid biosynthesis and UPR. NES: normalized enrichment score. FDR: false discovery rate. (E) Heatmap of DEGs in lipid synthesis and metabolism. (F) Heatmap of DEGs in UPR. (G) Protein expression of BiP, PERK, MVD, and IDI1 under different shear stress conditions. Data are expressed as the mean ± standard deviation from three or more independent experiments. Statistical significance was determined using an unpaired t-test, with * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. KEGG: Kyoto Encyclopedia of Genes and Genomes, GO: gene ontology, GSEA: gene set enrichment analysis.

Article Snippet: Antibodies: SLC7A11 (1:1000, ab175186, Abcam), GRP78/BIP (1:1000, 66574-1-Ig, proteintech), PERK/EIF2AK3 (1:1000, 24390-1-AP, proteintech), KLF6 (1:1000, 14716-1-AP, proteintech), MVD (1:1000, 15331-1-AP proteintech) IDI1 (1:1000, 11166-2-AP, proteintech), Goat Anti-Rabbit IgG-HRP (1:10,000, ASS1006-1, Biorigin), Goat Anti-Mouse IgG-HRP (1:10,000, ASS1007-1, Biorigin).

Techniques: RNA Sequencing, Expressing, Shear, Standard Deviation

Journal: bioRxiv

Article Title: Abnormal shear stress induces ferroptosis in endothelial cells via KLF6 downregulation

doi: 10.1101/2025.09.10.675441

Figure Lengend Snippet: KLF6 regulates the UPR and mevalonate pathway. (A) Heat map of the identified TF genes downregulated by LSS; (B) Predicted KLF6-binding motifs in promoters of PERK, HSPA5, MVD, IDI1 (JASPAR database; matrix ID: MA0472.1); (C) Protein expression of KLF6 under different shear stress conditions; (D) Validation of KLF6 overexpression by Western blot (OE: overexpression, NC: negative control); (E, F) KLF6-OE largely restores the expression of PERK, BiP, MVD, and IDI1 under LSS and HSS to levels seen under MSS. Data are expressed as the mean ± standard deviation from three or more independent experiments. Statistical significance was determined using an unpaired t-test, with * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Antibodies: SLC7A11 (1:1000, ab175186, Abcam), GRP78/BIP (1:1000, 66574-1-Ig, proteintech), PERK/EIF2AK3 (1:1000, 24390-1-AP, proteintech), KLF6 (1:1000, 14716-1-AP, proteintech), MVD (1:1000, 15331-1-AP proteintech) IDI1 (1:1000, 11166-2-AP, proteintech), Goat Anti-Rabbit IgG-HRP (1:10,000, ASS1006-1, Biorigin), Goat Anti-Mouse IgG-HRP (1:10,000, ASS1007-1, Biorigin).

Techniques: Binding Assay, Expressing, Shear, Biomarker Discovery, Over Expression, Western Blot, Negative Control, Standard Deviation

Journal: Frontiers in Nutrition

Article Title: Prevention of high-fat/high-sugar diet-induced type 2 diabetes mellitus-associated non-alcoholic fatty liver disease in rats with fermented and raw Rosa roxburghii Tratt (Cili) juice

doi: 10.3389/fnut.2025.1584551

Figure Lengend Snippet: Expression analyses of transcription or protein levels of selected genes. (A) Expression analyses of selected genes by qRT–PCR. The data represent the means ± SDs from six biological replicates with three technical replicates. (B–D) FCJ/RCJ upregulated ABCC3, IDI1, and APOA2 expression in the liver. FCJ and RCJ significantly upregulated hepatic ABCC3 (B) and IDI1 (C) expression, and RCJ significantly upregulated hepatic APOA2 (D) expression in T2DM rats. The data represent the means ± SDs from three biological replicates with three technical replicates. * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: The western blotting antibodies used included APOA2 (BM5624, BOSTER, China), IDI1 (A07892-1, BOSTER, China), ABCC3 (DF3874, Affinity, United States), β-tubulin (Solarbio, China), goat anti-rabbit (BS13278, Bioworld, United States), and goat anti-mouse (BS12478, Bioworld, United States) antibodies.

Techniques: Expressing, Quantitative RT-PCR