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Identification of hub genes and mRNAs-miRNAs co-expression networks. A Venn diagrams of the seven central genes screened by the 4 algorithms. B Display of the mRNA-miRNA network using cytokeratin. IL1B has five target miRNAs, ICAM1 has 14 target miRNAs, CCL2 has six target miRNAs, <t>TLR2</t> has five target miRNAs, and TLR4 has eleven target miRNAs. Red circles represent the mRNAs, blue circles represent miRNAs. MCC Maximum clustered centrality, EPC Edge Percolated Couponent, MNC Maximum neighborhood component
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DCA and LTA Treatment Regulate Macrophage Polarization in HFD-fed DEN Mice (A-C)The level of IL-6, IL-1β, and TNF-β in mice liver, n = 3. (D–G) The level of IL-6, IL-10, TGF-β, and TNF-α in liver of mice, n = 3. (H) The gene expression of Arg1, iNOS, CD206, TGR5, TLR2 in liver of mice, n = 3. (I) the protein level of p-NF-κB, p-STAT3, GPC-3 were detected by Western blot, β-actin is an internal parameter. (J) the quantified data of p-NF-κB, p-STAT3, GPC-3 of liver in Mice n = 3. Data are presented as means ± SD. ∗P < 0.05∗∗P < 0.01 ∗∗∗P < 0.001.

Journal: Metabolism Open

Article Title: Deoxycholic Acid and Lipoteichoic Acid cooperatively drive macrophage M2/M1 polarization via TGR5/STAT3 and TLR2/NF-κB to fuel HCC progression in obesity

doi: 10.1016/j.metop.2025.100420

Figure Lengend Snippet: DCA and LTA Treatment Regulate Macrophage Polarization in HFD-fed DEN Mice (A-C)The level of IL-6, IL-1β, and TNF-β in mice liver, n = 3. (D–G) The level of IL-6, IL-10, TGF-β, and TNF-α in liver of mice, n = 3. (H) The gene expression of Arg1, iNOS, CD206, TGR5, TLR2 in liver of mice, n = 3. (I) the protein level of p-NF-κB, p-STAT3, GPC-3 were detected by Western blot, β-actin is an internal parameter. (J) the quantified data of p-NF-κB, p-STAT3, GPC-3 of liver in Mice n = 3. Data are presented as means ± SD. ∗P < 0.05∗∗P < 0.01 ∗∗∗P < 0.001.

Article Snippet: Primary antibody concentrations were: p-NFκB p65 (ab76302, a dilution ratio of 1:1000, Abcam), p-STAT3(bs-1658R, a dilution ratio of 1:1000, Bioss), GPC-3 (30021-1-AP, a dilution ratio of 1:1000, Proteintech), β-actin (66009-1-Ig, a dilution ratio of 1:2000, Proteintech), 53BP1 (ab175933, a dilution ratio of 1:1000, abcam), TLR2 (ab209216, a dilution ratio of 1:1000, Proteintech), TGR5 (ab72608, a dilution ratio of 1:1000, abcam), E-cadherin (ABP0083, a dilution ratio of 1:1000, Abbkine), N-cadherin (22018-1-AP, a dilution ratio of 1:1000, Proteintech), GPC-3 (30021-1-AP, a dilution ratio of 1:1000, Proteintech).

Techniques: Gene Expression, Western Blot

DCA and LTA treatment promote polarization of Raw264.7 cells towards M2 and M1 types. (A–D) Representative immunofluorescence pictures and quantified data of iNOS and Arg1 in raw264.7 cell, Bar = 50 μm. n = 3. (E–J) The gene expression of TGF-β, IL-10, Arg1, TNF-α, IL-6, and iNOS in raw264.7 cell, n = 3. (K) Representative Western blot pictures and quantified data of TLR2, TGR5, p-NF-Κb, p-STAT3, in raw264.7 cell, β-actin is an internal parameter., n = 3. (P–S) The level of TNF-α, IL-6, IL-10, and TGF-βin supernatant of culture medium of raw264.7 cell, n = 3. Data are presented as means ± SD. ∗P < 0.05∗∗P < 0.01 ∗∗∗P < 0.001.

Journal: Metabolism Open

Article Title: Deoxycholic Acid and Lipoteichoic Acid cooperatively drive macrophage M2/M1 polarization via TGR5/STAT3 and TLR2/NF-κB to fuel HCC progression in obesity

doi: 10.1016/j.metop.2025.100420

Figure Lengend Snippet: DCA and LTA treatment promote polarization of Raw264.7 cells towards M2 and M1 types. (A–D) Representative immunofluorescence pictures and quantified data of iNOS and Arg1 in raw264.7 cell, Bar = 50 μm. n = 3. (E–J) The gene expression of TGF-β, IL-10, Arg1, TNF-α, IL-6, and iNOS in raw264.7 cell, n = 3. (K) Representative Western blot pictures and quantified data of TLR2, TGR5, p-NF-Κb, p-STAT3, in raw264.7 cell, β-actin is an internal parameter., n = 3. (P–S) The level of TNF-α, IL-6, IL-10, and TGF-βin supernatant of culture medium of raw264.7 cell, n = 3. Data are presented as means ± SD. ∗P < 0.05∗∗P < 0.01 ∗∗∗P < 0.001.

Article Snippet: Primary antibody concentrations were: p-NFκB p65 (ab76302, a dilution ratio of 1:1000, Abcam), p-STAT3(bs-1658R, a dilution ratio of 1:1000, Bioss), GPC-3 (30021-1-AP, a dilution ratio of 1:1000, Proteintech), β-actin (66009-1-Ig, a dilution ratio of 1:2000, Proteintech), 53BP1 (ab175933, a dilution ratio of 1:1000, abcam), TLR2 (ab209216, a dilution ratio of 1:1000, Proteintech), TGR5 (ab72608, a dilution ratio of 1:1000, abcam), E-cadherin (ABP0083, a dilution ratio of 1:1000, Abbkine), N-cadherin (22018-1-AP, a dilution ratio of 1:1000, Proteintech), GPC-3 (30021-1-AP, a dilution ratio of 1:1000, Proteintech).

Techniques: Immunofluorescence, Gene Expression, Western Blot

DCA and LTA promote hepatocellular carcinoma cell proliferation and migration by regulating the TGR5 and TLR2 pathways in RAW264.7 cells. (A–G) The gene expression of N-cadherin, Cdkn1a, Cdkn2a, TLR2, TGR5, and E-cadherin in co-cultured hepa1-6 cells. (H–O) Representative Western blot pictures and quantified data of E-cadherin,TLR2, TGR5, p-NF-Κb, GPC3, 53BP1, TLR2, TGR5 in co-cultured hepa1-6 cell, β-actin is an internal parameter., n = 3. (P,Q) Representative pictures and quantified data of migration of co-cultured hepa1-6 cells. (R,S) Representative pictures and quantified data of invasion of co-cultured hepa1-6 cells, Data are presented as means ± SD. ∗P < 0.05∗∗P < 0.01 ∗∗∗P < 0.001.

Journal: Metabolism Open

Article Title: Deoxycholic Acid and Lipoteichoic Acid cooperatively drive macrophage M2/M1 polarization via TGR5/STAT3 and TLR2/NF-κB to fuel HCC progression in obesity

doi: 10.1016/j.metop.2025.100420

Figure Lengend Snippet: DCA and LTA promote hepatocellular carcinoma cell proliferation and migration by regulating the TGR5 and TLR2 pathways in RAW264.7 cells. (A–G) The gene expression of N-cadherin, Cdkn1a, Cdkn2a, TLR2, TGR5, and E-cadherin in co-cultured hepa1-6 cells. (H–O) Representative Western blot pictures and quantified data of E-cadherin,TLR2, TGR5, p-NF-Κb, GPC3, 53BP1, TLR2, TGR5 in co-cultured hepa1-6 cell, β-actin is an internal parameter., n = 3. (P,Q) Representative pictures and quantified data of migration of co-cultured hepa1-6 cells. (R,S) Representative pictures and quantified data of invasion of co-cultured hepa1-6 cells, Data are presented as means ± SD. ∗P < 0.05∗∗P < 0.01 ∗∗∗P < 0.001.

Article Snippet: Primary antibody concentrations were: p-NFκB p65 (ab76302, a dilution ratio of 1:1000, Abcam), p-STAT3(bs-1658R, a dilution ratio of 1:1000, Bioss), GPC-3 (30021-1-AP, a dilution ratio of 1:1000, Proteintech), β-actin (66009-1-Ig, a dilution ratio of 1:2000, Proteintech), 53BP1 (ab175933, a dilution ratio of 1:1000, abcam), TLR2 (ab209216, a dilution ratio of 1:1000, Proteintech), TGR5 (ab72608, a dilution ratio of 1:1000, abcam), E-cadherin (ABP0083, a dilution ratio of 1:1000, Abbkine), N-cadherin (22018-1-AP, a dilution ratio of 1:1000, Proteintech), GPC-3 (30021-1-AP, a dilution ratio of 1:1000, Proteintech).

Techniques: Migration, Gene Expression, Cell Culture, Western Blot

Identification of hub genes and mRNAs-miRNAs co-expression networks. A Venn diagrams of the seven central genes screened by the 4 algorithms. B Display of the mRNA-miRNA network using cytokeratin. IL1B has five target miRNAs, ICAM1 has 14 target miRNAs, CCL2 has six target miRNAs, TLR2 has five target miRNAs, and TLR4 has eleven target miRNAs. Red circles represent the mRNAs, blue circles represent miRNAs. MCC Maximum clustered centrality, EPC Edge Percolated Couponent, MNC Maximum neighborhood component

Journal: Clinical and Experimental Medicine

Article Title: Integrated bioinformatics decoding of the MASLD ceRNA network reveals novel therapeutic targets

doi: 10.1007/s10238-025-01890-x

Figure Lengend Snippet: Identification of hub genes and mRNAs-miRNAs co-expression networks. A Venn diagrams of the seven central genes screened by the 4 algorithms. B Display of the mRNA-miRNA network using cytokeratin. IL1B has five target miRNAs, ICAM1 has 14 target miRNAs, CCL2 has six target miRNAs, TLR2 has five target miRNAs, and TLR4 has eleven target miRNAs. Red circles represent the mRNAs, blue circles represent miRNAs. MCC Maximum clustered centrality, EPC Edge Percolated Couponent, MNC Maximum neighborhood component

Article Snippet: IL-1β antibody (1:500, Cat# 16,806–1-AP, Proteintech); TLR2 antibody (1:500, Cat# 66,625–1-Ig, Proteintech); TLR4 antibody (1:500, Cat# 19,811–1-AP, Proteintech);β-Tubulin antibody (1:20,000, Cat# 66,240–1-Ig, Proteintech).

Techniques: Expressing

Hub genes were validated and ROC curve analyzed using another human Metabolic dysfunction-associated steatotic liver disease dataset. A ICAM1, IL1B, TLR2 and TLR4 expression was up-regulated in patients with Metabolic dysfunction-associated steatotic liver disease compared to controls. *: p < 0.001.0 1, **: p < 0.0 5. B ICAM1 and IL1B had the highest AUC values (1.000), followed by ITGAM (AUC value: 0.991), followed by TLR2 (AUC value: 0.979), CCL2 (AUC value: 0.829) and lastly TLR4 (AUC value: 0.667). ROC Receiver Operating Characteristic, AUC Area under the curve

Journal: Clinical and Experimental Medicine

Article Title: Integrated bioinformatics decoding of the MASLD ceRNA network reveals novel therapeutic targets

doi: 10.1007/s10238-025-01890-x

Figure Lengend Snippet: Hub genes were validated and ROC curve analyzed using another human Metabolic dysfunction-associated steatotic liver disease dataset. A ICAM1, IL1B, TLR2 and TLR4 expression was up-regulated in patients with Metabolic dysfunction-associated steatotic liver disease compared to controls. *: p < 0.001.0 1, **: p < 0.0 5. B ICAM1 and IL1B had the highest AUC values (1.000), followed by ITGAM (AUC value: 0.991), followed by TLR2 (AUC value: 0.979), CCL2 (AUC value: 0.829) and lastly TLR4 (AUC value: 0.667). ROC Receiver Operating Characteristic, AUC Area under the curve

Article Snippet: IL-1β antibody (1:500, Cat# 16,806–1-AP, Proteintech); TLR2 antibody (1:500, Cat# 66,625–1-Ig, Proteintech); TLR4 antibody (1:500, Cat# 19,811–1-AP, Proteintech);β-Tubulin antibody (1:20,000, Cat# 66,240–1-Ig, Proteintech).

Techniques: Expressing

Three CERNA networks, TLR2, TLR4 and IL1B, and their potential RNA regulatory pathways. A–C CERNA network of TLR2, TLR4 and IL1B. Red indicates central genes, green indicates miRNAs, and blue indicates IncRNAs

Journal: Clinical and Experimental Medicine

Article Title: Integrated bioinformatics decoding of the MASLD ceRNA network reveals novel therapeutic targets

doi: 10.1007/s10238-025-01890-x

Figure Lengend Snippet: Three CERNA networks, TLR2, TLR4 and IL1B, and their potential RNA regulatory pathways. A–C CERNA network of TLR2, TLR4 and IL1B. Red indicates central genes, green indicates miRNAs, and blue indicates IncRNAs

Article Snippet: IL-1β antibody (1:500, Cat# 16,806–1-AP, Proteintech); TLR2 antibody (1:500, Cat# 66,625–1-Ig, Proteintech); TLR4 antibody (1:500, Cat# 19,811–1-AP, Proteintech);β-Tubulin antibody (1:20,000, Cat# 66,240–1-Ig, Proteintech).

Techniques:

Expression Levels of IL-1B, TLR2, and TLR4 in Normal Liver Tissues and Liver Tissues of MASLD Patients as Revealed by Multiplexed Immunohistochemistry Fluorescence and Western Blot. A–D Multiplexed immunohistochemical fluorescence detection of pivotal gene expression levels in normal liver tissues and tissues with Metabolic dysfunction-associated steatotic liver disease. E–H Western Blot Detection of hub genes Protein Expression in Normal Liver Tissue and MASLD Tissue. MASLD: Metabolic dysfunction-associated steatotic liver disease. ( n = 3 mice/group; scale bar: 50 µm). Statistical significance by unpaired t test * p < 0.05, **** p < 0.0001

Journal: Clinical and Experimental Medicine

Article Title: Integrated bioinformatics decoding of the MASLD ceRNA network reveals novel therapeutic targets

doi: 10.1007/s10238-025-01890-x

Figure Lengend Snippet: Expression Levels of IL-1B, TLR2, and TLR4 in Normal Liver Tissues and Liver Tissues of MASLD Patients as Revealed by Multiplexed Immunohistochemistry Fluorescence and Western Blot. A–D Multiplexed immunohistochemical fluorescence detection of pivotal gene expression levels in normal liver tissues and tissues with Metabolic dysfunction-associated steatotic liver disease. E–H Western Blot Detection of hub genes Protein Expression in Normal Liver Tissue and MASLD Tissue. MASLD: Metabolic dysfunction-associated steatotic liver disease. ( n = 3 mice/group; scale bar: 50 µm). Statistical significance by unpaired t test * p < 0.05, **** p < 0.0001

Article Snippet: IL-1β antibody (1:500, Cat# 16,806–1-AP, Proteintech); TLR2 antibody (1:500, Cat# 66,625–1-Ig, Proteintech); TLR4 antibody (1:500, Cat# 19,811–1-AP, Proteintech);β-Tubulin antibody (1:20,000, Cat# 66,240–1-Ig, Proteintech).

Techniques: Expressing, Immunohistochemistry, Fluorescence, Western Blot, Immunohistochemical staining, Gene Expression