Journal: Chinese Medicine

Article Title: Panduratin A from Boesenbergia rotunda suppresses hepatitis B virus by targeting HNF1α and synergizing with antiviral agents

doi: 10.1186/s13020-025-01285-w

Figure Lengend Snippet: Boesenbergia rotunda extract and panduratin A reduced HNF1α expression at both mRNA and protein levels. imHC cells were transfected with 0–2 µg of pHBV 1.3-mer WT. On day 3, HNF1α ( A ) and HNF4α ( B ) mRNA levels were measured by real-time qPCR. The mRNA expression of liver-enriched transcription factors, including HNF1α, HNF4α, PPARα, and C/EBPα, was analyzed after treatment with 0–10 µM panduratin A ( C ). The effects of B. rotunda extract and panduratin A on HNF1α ( D ) and HNF4α ( E ) protein expression were assessed by Western Blot. HBV-infected d-imHC cells were treated with 0–1.25 µM panduratin A, and HNF1α and HNF4α protein levels were detected by Western Blot ( F ). Western Blot quantification was performed using ImageJ software, with normalization to GAPDH

Article Snippet: To confirm the critical role of HNF1α in the anti-HBV activity of panduratin A, HNF1α expression was either silenced or ectopically expressed in hepatocytes using HNF1α-shRNA plasmid (sc-35567-SH, Santa Cruz Biotechnology) or an HNF1α plasmid (Addgene plasmid #31104), respectively, to assess the loss- and gain-of-function effects.

Techniques: Expressing, Transfection, Western Blot, Infection, Software

Journal: Chinese Medicine

Article Title: Panduratin A from Boesenbergia rotunda suppresses hepatitis B virus by targeting HNF1α and synergizing with antiviral agents

doi: 10.1186/s13020-025-01285-w

Figure Lengend Snippet: The anti-HBV activity of panduratin A is mediated through HNF1α and is impaired by HNF1α knockdown but restored by HNF1α rescue. imHC cells were transfected with sh-Ctrl, sh-HNF1α or HNF1α plasmids for 4 h before treatment with 0–10 µM panduratin A. On day 3, cell viability was assessed using the MTT assay ( A ). The mRNA expression of liver-enriched transcription factors (HNF1α, HNF4α, C/EBPα, and FoxO4) was analyzed in sh-Ctrl and HNF1α-knockdown imHCs ( B ). HNF1α-knockdown imHCs were treated with panduratin A, and the protein levels of HNF1α, HNF4α, and GAPDH were detected by Western Blot ( C ). Nuclear localization of HNF1α was analyzed in imHCs transfected with sh-HNF1α or sh-Ctrl following treatment with 5 µM panduratin A or 0.1% DMSO. Scale bar, 50 µm ( D ). HBV RNA levels were measured in infected imHCs transfected with sh-Ctrl, sh-HNF1α, or an HNF1α expression plasmid following treatment with 0–10 µM panduratin A ( E ), and HBsAg levels were quantified by ELISA ( F ). HBV promoter activities were evaluated in HNF1α-knockdown cells with or without panduratin A treatment; pCMV and pLuc served as negative and positive controls, respectively ( G ). Immunofluorescence analysis detected HBV core protein (green) and nuclei (DAPI, blue) in sh-HNF1α and HNF1α-transfected imHCs with or without 5 µM panduratin A treatment ( H ). HNF1α expression, HBV RNA levels, and viral load were measured in wild-type, HNF1α-knockdown, and HNF1α-knockdown cells with ectopic HNF1α expression, with or without panduratin A treatment ( I ). Scale bar, 50 µm. Data were presented as mean ± SD. * or a, ** or b, *** or c, and **** or d indicate statistical significance at p value < 0.05, < 0.01, < 0.001, and < 0.0001, respectively

Article Snippet: To confirm the critical role of HNF1α in the anti-HBV activity of panduratin A, HNF1α expression was either silenced or ectopically expressed in hepatocytes using HNF1α-shRNA plasmid (sc-35567-SH, Santa Cruz Biotechnology) or an HNF1α plasmid (Addgene plasmid #31104), respectively, to assess the loss- and gain-of-function effects.

Techniques: Activity Assay, Knockdown, Transfection, MTT Assay, Expressing, Western Blot, Infection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Chinese Medicine

Article Title: Panduratin A from Boesenbergia rotunda suppresses hepatitis B virus by targeting HNF1α and synergizing with antiviral agents

doi: 10.1186/s13020-025-01285-w

Figure Lengend Snippet: Interaction networks of HNF1α, HNF4α, PPARα, and C/EBPα based on GeneMANIA. Gene interaction analysis was conducted using GeneMANIA to explore potential mechanisms underlying the anti-HBV effects of panduratin A. The analysis visualizes interaction types (indicated by different colors), interaction strengths (represented by edge thickness), multiple edges between nodes, and protein scores (reflected by node sizes) ( A ). A schematic diagram illustrating the proposed mechanism by which panduratin A inhibits HBV replication in hepatocytes. Panduratin A suppresses HNF1α, which in turn regulates the HBV promoter. Created in BioRender, https://BioRender.com/q22t488 ( B )

Article Snippet: To confirm the critical role of HNF1α in the anti-HBV activity of panduratin A, HNF1α expression was either silenced or ectopically expressed in hepatocytes using HNF1α-shRNA plasmid (sc-35567-SH, Santa Cruz Biotechnology) or an HNF1α plasmid (Addgene plasmid #31104), respectively, to assess the loss- and gain-of-function effects.

Techniques:

Journal: International Journal of Medical Sciences

Article Title: Missense and Intronic Variants in HNF1A Affect Prostate Cancer Aggressiveness in Patients with Biochemical Recurrence

doi: 10.7150/ijms.127638

Figure Lengend Snippet: Correlation between HNF1A expression and clinical features in patients with PCa based on TCGA-PRAD data. (A) HNF1A expression levels in unpaired normal and tumor tissues in the TCGA-PRAD dataset are presented. (B) HNF1A expression levels were analyzed in 52 matched PCa tissues and their corresponding normal tissues. (C-E) HNF1A expression levels in PCa from TCGA-PRAD were compared based on the Gleason scores (C), pathological T stages (D), and distant metastasis (E). (F) Kaplan-Meier survival curves were used to illustrate OS of patients with high and low HNF1A expression.

Article Snippet: We conducted genotyped rs2464196 (assay ID: C___1263617_10), rs1169288 (assay ID: C___7474231_10), rs735396 (assay ID: C___1263608_1_), and rs1169286 (assay ID: C___1263544_20) assays by using TaqMan SNP Genotyping Assay on an ABI StepOnePlus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Expressing

Journal: International Journal of Medical Sciences

Article Title: Missense and Intronic Variants in HNF1A Affect Prostate Cancer Aggressiveness in Patients with Biochemical Recurrence

doi: 10.7150/ijms.127638

Figure Lengend Snippet: HNF1A-associated pathways in patients with PCa. The horizontal bar plot displays pathways identified in the Hallmark database that are correlated with HNF1A expression. Pathways positively and negatively associated with HNF1A are displayed in red and blue, respectively. The x -axis indicates normalized enrichment scores (NESs).

Article Snippet: We conducted genotyped rs2464196 (assay ID: C___1263617_10), rs1169288 (assay ID: C___7474231_10), rs735396 (assay ID: C___1263608_1_), and rs1169286 (assay ID: C___1263544_20) assays by using TaqMan SNP Genotyping Assay on an ABI StepOnePlus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: LTH-sEV promotes LSEC angiogenesis by regulating HNF1α/Ang-2 axis. ( A ). Diagram of a database cross to confirm transcription factor HNF1α; ( B ) qRT-PCR analysis of Ang-2 mRNA expression in LSECs transfected with pCtr or different doses of pHNF1α, n = 6; ( C ) Western blot analysis of HNF1α and Ang-2 protein expression in LSECs transfected with pCtr or different doses of pHNF1α, n = 3; ( D ) qRT-PCR analysis of Ang-2 mRNA expression in LSECs transfected with siCtr, siHNF1α-1or siHNF1α-2, n = 6; ( E ) Western blot analysis of HNF1α and Ang-2 protein expression in LSECs transfected with siCtr, siHNF1α-1, or siHNF1α-2, n = 3; ( F and G ) JASPAR database predicts the binding sites of HNF1α and Ang-2 and constructs the truncation; ( H ) Ang-2 promoter activity in LSECs transfected with different doses of pHNF1α or pCtr and pAng-2-2000; Ang-2 promoter activity in LSECs transfected with siCtr, siHNF1α-1or siHNF1α-2 and pAng-2-2000, n = 4; ( I ) Ang-2 promoter activity in 293T and LSECs transfected with pHNF1α and pAng-2, n = 4; ( J ) ChIP-PCR analysis of the binding area of HNF1α to Ang-2 in 293T and LSECs transfected with pCtr or pHNF1α, n = 4; ( K and L ) qRT-PCR and Western blot of HNF1α and Ang-2 protein expression in normal LSECs or LTH-sEV treated LSECs transfected with siCtr or siHNF1α, n = 6; ( M and N ) Fibrin gel bead assay and tube formation assay in LSECs or LTH-sEV-treated LSECs transfected with siCtr or siHNF1α, scale bar = 50/200 μm, n = 5; Compared with the siCtr group, ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Quantitative RT-PCR, Expressing, Transfection, Western Blot, Binding Assay, Construct, Activity Assay, Tube Formation Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: LTH-sEV transports OGT into LSEC and liver to promote HNF1α O-GlcNAcylation. ( A ) qRT-PCR of HNF1α mRNA expression in LSECs treated with PBS, LTH-sEV (200, 400 μg/mL), n = 6; Compared with the NC group, ns, no significance; ( B ) IP-MS analysis of the protein binding to HNF1α in LSECs treated with PBS, LTH-sEV (400 μg/mL); ( C ) Co-IP of cell lysates from LSECs using antibodies against HNF1α; ( D ) Immunofluorescence images of the co-localization of OGT and HNF1α in LSECs, scale bar, 20 μm; ( E ) Western blot analysis of OGT expression in LTH-sEV; ( F ) qRT-PCR of OGT mRNA expression in LSECs treated with PBS, LTH-sEV (200, 400 μg/mL), n = 6; Compared with the PBS group, ns, no significance; ( G ) Western blot analysis of OGT, OGA, and O-GlcNac expression in LSECs treated with PBS, LTH-sEV (200, 400 μg/mL), n = 3; ( H ) Immunofluorescence images of co-localization of OGT and DiR-labeled LTH-sEV in LSEC, scale bar = 20 μm; ( I ) Western Blot analysis of OGT, OGA, and O-GlcNac protein expression in mouse liver. n = 3; ( J ) Immunofluorescence images of co-localization of OGT and CD63 in mouse liver, scale bar = 50 μm; ( K ) Immunohistochemistry images of OGT and O-GlcNac, scale bar = 50 μm, n = 6; ( L ) Co-IP of cell lysates from 293T transfected with V5-HNF1α using antibodies against V5-HNF1α; ( M ) Co-IP of cell lysates from LSECs using antibodies against HNF1α; ( N ) Co-IP of cell lysates from 293T transfected with V5-HNF1α and treated with PBS, LTH-sEV using antibodies against V5-HNF1α; ( O ) Co-IP of cell lysates from LSEC treated with PBS, LTH-sEV using antibodies against HNF1α.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Quantitative RT-PCR, Expressing, Protein-Protein interactions, Protein Binding, Co-Immunoprecipitation Assay, Immunofluorescence, Western Blot, Labeling, Immunohistochemistry, Transfection

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: OGT promotes Ang-2 expression and LSEC angiogenesis through HNF1α O-GlcNAcylation at Ser471 site. ( A and B ) Co-IP of cell lysates from 293T transfected with V5-HNF1α and His-OGT or pCtr, siCtr, or siOGT using antibodies against V5-HNF1α; ( C ) Immunofluorescence images of HNF1α in LSEC transfected with shOGT or shCtr, scale bar = 10 μm; ( D ) YinOYang 1.2 website predicts O-GlcNAc sites of HNF1α; ( E ) Co-IP of cell lysates from 293T transfected with pCtr or various V5-HNF1α mutants using antibodies against V5-HNF1α; ( F ) Co-IP of cell lysates from 293T transfected with pCtr or various V5-HNF1α mutants and His-OGT using antibodies against V5-HNF1α; ( G ) qRT-PCR analysis of Ang-2 mRNA expression in 293T and LSECs transfected with pCtr and V5-HNF1α mutant; Compared with V5-HNF1α-WT group, ∗∗ P < .01; ∗∗∗ P < .001; ns, no significance; ( H ) Immunofluorescence images of V5 in LSEC transfected with pCtr, V5-HNF1α-WT or V5-HNF1α-S471A, scale bar = 10 μm; ( I–L ) Normal LSEC and OGT knocked down LSECs transfected with pCtr,V5-HNF1α-WT or V5-HNF1α-S471A; ( I ) qRT-PCR analysis of Ang-2 mRNA expression, n = 6; ( J ) Western blot analysis of Ang-2 protein expression, n = 3; ( K and L ) Fibrin gel bead assay and tube formation assay of LSEC, scale bar = 50/200 μm, n = 5. Compared with the pCtr group, ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ns, no significance.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Expressing, Co-Immunoprecipitation Assay, Transfection, Immunofluorescence, Quantitative RT-PCR, Mutagenesis, Western Blot, Tube Formation Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: OGT enriched MASLD-sEV promotes LSEC angiogenesis. ( A and B ) qRT-PCR and Western blot analysis of OGT expression in LSECs treated with Healthy-sEV and MASLD-sEV; ( C ) Immunofluorescence images of co-localization of OGT and DiR-labeled Healthy-sEV and MASLD-sEV, scale bar = 20 μm; ( D ) Co-IP of cell lysates from 293T transfected with V5-HNF1α and treated with Healthy-sEV and MASLD-sEV using antibodies against V5-HNF1α; ( E and F ) qRT-PCR and Western blot analysis of Ang-2 expression in LSEC treated with Healthy-sEV and MASLD-sEV, n = 6 or 3; ( G and H ) Fibrin gel bead assay and tube formation assay in LSECs treated with PBS, Healthy-sEV and MASLD-sEV, scale bar = 50/200 μm, n = 5; Compared with Healthy-sEV or NC group, ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ns, no significance.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Labeling, Co-Immunoprecipitation Assay, Transfection, Tube Formation Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: BAGN inhibits LSEC capillarization and alleviates MASLD progression. ( A ) H&E staining of liver, heart, spleen, lung, and kidney in HFD mice treated with DMSO or BAGN (0.5 mg, 1.0 mg, 2.0 mg), scale bar = 50 μm; ( B ) Western blot analysis of O-GlcNac, OGT, and Ang-2 protein expression in HFD mice, n = 3; ( C ) Immunofluorescence images of HNF1α and O-GlcNac, scale bar = 20 μm; ( D ) SEM images and porosity of liver in each group, scale bar = 500 nm, n = 6; Immunohistochemistry images of CD31 and Ang-2, scale bar = 50 μm, n = 6; ( E ) Serum ALT, AST, and TG expression in each group, n = 6; ( F ) Serum IL-1β, IL-6, and TNFα expression in each group, n = 6; ( G ) Oil red O staining, immunohistochemistry images of α-SMA, Sirius red staining in each group, scale bar = 20/100/50 μm, n = 6; Compared with HFD group, ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Staining, Western Blot, Expressing, Immunofluorescence, Immunohistochemistry

Journal: International Journal of Medical Sciences

Article Title: Missense and Intronic Variants in HNF1A Affect Prostate Cancer Aggressiveness in Patients with Biochemical Recurrence

doi: 10.7150/ijms.127638

Figure Lengend Snippet: Correlation between HNF1A expression and clinical features in patients with PCa based on TCGA-PRAD data. (A) HNF1A expression levels in unpaired normal and tumor tissues in the TCGA-PRAD dataset are presented. (B) HNF1A expression levels were analyzed in 52 matched PCa tissues and their corresponding normal tissues. (C-E) HNF1A expression levels in PCa from TCGA-PRAD were compared based on the Gleason scores (C), pathological T stages (D), and distant metastasis (E). (F) Kaplan-Meier survival curves were used to illustrate OS of patients with high and low HNF1A expression.

Article Snippet: We conducted genotyped rs2464196 (assay ID: C___1263617_10), rs1169288 (assay ID: C___7474231_10), rs735396 (assay ID: C___1263608_1_), and rs1169286 (assay ID: C___1263544_20) assays by using TaqMan SNP Genotyping Assay on an ABI StepOnePlus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Expressing

Journal: International Journal of Medical Sciences

Article Title: Missense and Intronic Variants in HNF1A Affect Prostate Cancer Aggressiveness in Patients with Biochemical Recurrence

doi: 10.7150/ijms.127638

Figure Lengend Snippet: HNF1A-associated pathways in patients with PCa. The horizontal bar plot displays pathways identified in the Hallmark database that are correlated with HNF1A expression. Pathways positively and negatively associated with HNF1A are displayed in red and blue, respectively. The x -axis indicates normalized enrichment scores (NESs).

Article Snippet: We conducted genotyped rs2464196 (assay ID: C___1263617_10), rs1169288 (assay ID: C___7474231_10), rs735396 (assay ID: C___1263608_1_), and rs1169286 (assay ID: C___1263544_20) assays by using TaqMan SNP Genotyping Assay on an ABI StepOnePlus Real-Time PCR System (Thermo Fisher Scientific).

Techniques: Expressing