primary antibody incubation against tnf α  (Proteintech)

Journal: Frontiers in Nutrition

Article Title: Lactobacillus acidophilus alleviates slow transit constipation by modulating 5-HT pathway and gut microbial composition

doi: 10.3389/fnut.2026.1775405

Figure Lengend Snippet: Lactobacillus acidophilus modulates the expression of inflammatory cytokines. (A) Immunohistochemical analysis of TNF-α protein expression in mouse colon tissue. (B) RT-qPCR analysis of TNF-α mRNA expression in colonic tissues. (C) RT-qPCR analysis of IL-1β mRNA expression in colonic tissues. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibody incubation against TNF-α (Proteintech, Wuhan, China) (60291-1-IG) (dilution 1:1000) was performed overnight at 4 °C.

Techniques: Expressing, Immunohistochemical staining, Quantitative RT-PCR

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: Primary antibodies against tumor necrosis factor-α (TNF-α, #11948), interleukin-6 (IL-6, #12912), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, #2118S), Bax (#2772S), Bcl-2 (#3498S), and hemagglutinin (HA, #3724S) were purchased from Cell Signaling Technology (Shanghai, 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) Volcano plot analysis showing differentially expressed genes between the MgIG group (ALD+MgIG) and the A-Ctrl (ALD only) group after RNA-seq in liver of mice. (B) The top 5 regulated pathways (both up- and down-regulated) and the top 3 genes within each pathway are shown. (C) Volcano plot analysis showing differentially expressed genes between the A-Ctrl group and the Ctrl group after RNA-seq in liver of mice. (D) Changes in AML-12 cell viability, apoptosis, and Nile Red staining signals after ethanol/PA treatment, with or without co-treatment with different MgIG doses (0, 0.1, 0.25, 0.5, and 1 mg/ml) (n = 4). (E) Representative images of Nile Red staining in AML-12 cells treated with ethanol/PA, with or without co-treatment with different MgIG doses. (F) Changes in mRNA expression of lipid metabolism genes ( Acc1 and Scd1 ) and systemic inflammation markers ( Tnf-α and Il-6 ) in AML-12 cells treated with ethanol/PA, with or without co-treatment with different MgIG doses. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bar: 20 μm.

Article Snippet: Primary antibodies against tumor necrosis factor-α (TNF-α, #11948), interleukin-6 (IL-6, #12912), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, #2118S), Bax (#2772S), Bcl-2 (#3498S), and hemagglutinin (HA, #3724S) were purchased from Cell Signaling Technology (Shanghai, China).

Techniques: RNA Sequencing, Staining, 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) 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: Primary antibodies against tumor necrosis factor-α (TNF-α, #11948), interleukin-6 (IL-6, #12912), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, #2118S), Bax (#2772S), Bcl-2 (#3498S), and hemagglutinin (HA, #3724S) were purchased from Cell Signaling Technology (Shanghai, 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: Primary antibodies against tumor necrosis factor-α (TNF-α, #11948), interleukin-6 (IL-6, #12912), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, #2118S), Bax (#2772S), Bcl-2 (#3498S), and hemagglutinin (HA, #3724S) were purchased from Cell Signaling Technology (Shanghai, 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-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: Primary antibodies against tumor necrosis factor-α (TNF-α, #11948), interleukin-6 (IL-6, #12912), glyceraldehyde-3-phosphate dehydrogenase (GAPDH, #2118S), Bax (#2772S), Bcl-2 (#3498S), and hemagglutinin (HA, #3724S) were purchased from Cell Signaling Technology (Shanghai, China).

Techniques: Staining, Knockdown, Over Expression, Activity Assay

Journal: Journal of Inflammation Research

Article Title: Dual Effects of Sodium Houttuyfonate Against Acute Rhinosinusitis: Antibacterial Action and Epithelial Barrier Repair Through the p38 MAPK/ERK Pathway

doi: 10.2147/JIR.S539966

Figure Lengend Snippet: SH reduced inflammatory marker expression in ARS rat nasal mucosa. ( A ) Representative immunohistochemical staining of TNF-α (upper) and ICAM-1 (lower) in sinonasal mucosa across six groups (Control, Model, SH-D, SH-Z, SH-G, and AMC+ELAP-EC). ( B ) Quantification of TNF-α positive area (%). ( C ) Quantification of ICAM-1 positive area (%). *Data are presented as mean ± SD. (n = 3, ***p < 0.001 vs Control; #p < 0.05, ###p < 0.01 vs Model).

Article Snippet: Primary antibodies against TNF-α (Bioss, Catalog No. BS-10802R) and ICAM-1 (Proteintech, Catalog No. 16174-1-AP) were used for immunohistochemical analysis.

Techniques: Marker, Expressing, Immunohistochemical staining, Staining, Control

Journal: Poultry Science

Article Title: Music intervention mitigates LPS-induced gut barrier disruption and immune stress in broilers via TLR4/NF-κB regulation

doi: 10.1016/j.psj.2025.105189

Figure Lengend Snippet: Music has potential anti-inflammatory effects and can effectively alleviate inflammation in the jejunum and ileum by inhibiting the LPS-triggered TLR4/NF-κB signaling pathway. (A) The figure shows the relative mRNA expression levels of the TLR4/NF-κB inflammatory signaling pathway and related inflammatory factors in the jejunum and ileum of broilers. (B) The figure presents the immunoblot images and corresponding statistical results of TLR4, IKB-α, NF-κB p65, and TNF-α proteins in the jejunum and ileum of broilers. (C) The figure shows the measurement results of IL-1β, IL-6, and IL-10 levels in the jejunum and ileum of broilers across different groups. Differences marked with lowercase letters denote a significant difference ( P < 0.05), while the same letter indicates no significant difference ( P > 0.05).

Article Snippet: The membrane was then blocked in 5 % skim milk at 37°C for 2 h. After blocking, the membrane was incubated with diluted primary antibodies against TLR4, IKB-α, NF-κB, and TNF-α (1:1000, Abmart, China) at 4°C for 12 h. The specific antibodies used are listed in STable 6 .

Techniques: Expressing, Western Blot