Journal: Acta Cirúrgica Brasileira

Article Title: Hypofractionated radiotherapy followed by rhGM-CSF enhances immunogenic cell death in a murine model of pancreatic cancer

doi: 10.1590/acb407625

Figure Lengend Snippet: The workflow of treatment in pancreatic cancer in mice.

Article Snippet: The tumor sections were washed three times with PBS and then incubated in 3% peroxide for 3 min, followed by incubation in 5% bovine serum albumin (BSA) at room temperature for 30 min. Primary antibodies against hypoxia-inducible factor-1α (HIF-1α) (20960-1-AP, Proteintech), hypoxia-inducible factor-2α (HIF-2α) (PA1-16510, Thermo Fisher Scientific), soluble vascular endothelial growth factor (VEGF) receptor-1 (sVEGFR-1) (36-1100, Thermo Fisher Scientific), and VEGF (19003-1-AP, Proteintech) were used to incubate with tumor sections overnight at 4°C.

Techniques:

Journal: Acta Cirúrgica Brasileira

Article Title: Hypofractionated radiotherapy followed by rhGM-CSF enhances immunogenic cell death in a murine model of pancreatic cancer

doi: 10.1590/acb407625

Figure Lengend Snippet: rhGM-CSF combined with HFRT inhibits the angiogenesis-related factors expression in PC tumors. Tumors were treated with rhGM-CSF and HFRT, respectively, or in combination (HFRT followed by rhGM-CSF treatment, HFRT & rhGM-CSF treatment concurrently, and rhGM-CSF treatment followed by HFRT). ( a and e ) HIF-1α, ( b and f ) HIF-2α, ( c and g ) sVEGFR-1, ( d and h ) VEGF were measured by immunohistochemistry assay.

Article Snippet: The tumor sections were washed three times with PBS and then incubated in 3% peroxide for 3 min, followed by incubation in 5% bovine serum albumin (BSA) at room temperature for 30 min. Primary antibodies against hypoxia-inducible factor-1α (HIF-1α) (20960-1-AP, Proteintech), hypoxia-inducible factor-2α (HIF-2α) (PA1-16510, Thermo Fisher Scientific), soluble vascular endothelial growth factor (VEGF) receptor-1 (sVEGFR-1) (36-1100, Thermo Fisher Scientific), and VEGF (19003-1-AP, Proteintech) were used to incubate with tumor sections overnight at 4°C.

Techniques: Expressing, Immunohistochemistry

Journal: bioRxiv

Article Title: HIF1α-dependent induction of the T-Type calcium channel CaV3.2 mediates hypoxia-induced neuronal hyperexcitability

doi: 10.1101/2025.10.13.682038

Figure Lengend Snippet: NS20Y cells transfected with HIF1α show a significant upregulation of CaV3.2 in the (A) fluorescent reporter assay (Two-sided T Test, p = 0.013) (B) as well as in the dual Luciferase reporter assay (one-sample T Test, p = 0.037).

Article Snippet: Primary antibodies against HIF1α (Proteintech #20960-1-AP) were used in a concentration of 1:1000 in fish block over night at 4 °C.

Techniques: Transfection, Reporter Assay, Luciferase

Journal: bioRxiv

Article Title: HIF1α-dependent induction of the T-Type calcium channel CaV3.2 mediates hypoxia-induced neuronal hyperexcitability

doi: 10.1101/2025.10.13.682038

Figure Lengend Snippet: (A) primary neurons transduced with HIF1α show a significant upregulation of CaV3.2 in the Dual Luciferase reporter assay (one-sample T-Test, p = 0.016). (B) Primary neurons transduced with AAV-hSyn-HIF1α show a significantly increased WMFR compared to primary neurons transduced with a control virus (AAV-hSyn-GFP, two-way ANOVA HIF+/HIF– : F = 4.17, p = 0.049). (C) shows representative images of recordings of AAV-hSyn-GFP controls and AAV-hSyn-HIF1α with activity of every electrode shown in the left panel and bursting behavior on the right panel. (D) Artificial neural network analysis confirmed more hypoxia-like behavior of cells transduced with AAV-hSyn-HIF1α compared to control virus.

Article Snippet: Primary antibodies against HIF1α (Proteintech #20960-1-AP) were used in a concentration of 1:1000 in fish block over night at 4 °C.

Techniques: Transduction, Luciferase, Reporter Assay, Control, Virus, Activity Assay

Journal: bioRxiv

Article Title: HIF1α-dependent induction of the T-Type calcium channel CaV3.2 mediates hypoxia-induced neuronal hyperexcitability

doi: 10.1101/2025.10.13.682038

Figure Lengend Snippet: In murine OTCs (A) HIF1α shows a significant upregulation upon incubation in hypoxia in qPCR analysis compared to OTCs kept at normoxic conditions (two-sided paired T-Test, p = 0.05). (B) qPCR for Cacna1h revealed a drastic upregulation upon hypoxia compared to OTCs incubated in ambient oxygen concentration (two-sided paired T-Test, p = 0.009). (C) Human OTCs showed a significant upregulation of HIF1α upon exposure to hypoxia compared to OTCs kept at normoxic condition in western blot analysis (two-sided paired T-test, p = 0.04). Representative images of bands used for quantification are shown below (Ponceau S staining for WPN and HIF1α band at 120 kDa). (D) Analysis immunofluorescent stainings of CaV3.2 revealed a higher area fraction of CaV3.2 upon hypoxia compared to slices kept at normoxic conditions (two-sided paired T-Test, p = 0.03). (E) Representative images of immunofluorescent stainings of NeuN positive cells (neurons) and CaV3.2 staining. In normoxic conditions very little CaV3.2 expression is present whereas upregulation of CaV3.2 is visible on the apical dendrite of shown neuron. Scale bar corresponds to 10 µm.

Article Snippet: Primary antibodies against HIF1α (Proteintech #20960-1-AP) were used in a concentration of 1:1000 in fish block over night at 4 °C.

Techniques: Incubation, Concentration Assay, Western Blot, Staining, Expressing

Journal: bioRxiv

Article Title: HIF1α-dependent induction of the T-Type calcium channel CaV3.2 mediates hypoxia-induced neuronal hyperexcitability

doi: 10.1101/2025.10.13.682038

Figure Lengend Snippet: (A) Primary neurons exposed to the OGD/R model showed a significantly increased WMFR compared to neurons kept at normoxic conditions throughout the experiment (two-way ANOVA hypoxia × normoxia: F = 7.66, p = 0.009). (B) shows representative images of recordings of normoxic controls and hypoxia exposed OGD/R cells with activity of every electrode shown in the left panel and bursting behavior on the right panel. (C) Artificial neural network analysis confirmed similar behavior of hypoxia-exposed neurons to HIF+ (AAV-hSyn-HIF1α) cells, whereas neurons kept at normoxic conditions behaved rather like HIF-(AAV-hSyn-GFP) cells.

Article Snippet: Primary antibodies against HIF1α (Proteintech #20960-1-AP) were used in a concentration of 1:1000 in fish block over night at 4 °C.

Techniques: Activity Assay

Journal: Drug Design, Development and Therapy

Article Title: Cinobufagin Inhibits Invasion and Migration of Non-Small Cell Lung Cancer via Regulating Glucose Metabolism Reprogramming in Tumor-Associated Macrophages

doi: 10.2147/DDDT.S531190

Figure Lengend Snippet: Effect of CB on Genes Related to M2 Macrophage Polarization. ( A – C ) Real-time quantitative PCR analysis of mRNA levels for CD206, Arg-1, and IL-10; n=3; ** P <0.01 compared to M0; ## P <0.01 compared to M2.

Article Snippet: Primary antibodies anti-HIF-1α (1:200; Cat# 66730-1-Ig; Proteintech) or anti-PD-L1 (1:100; Cat# 17952-1-AP; Proteintech) were added followed by overnight incubation at 4°C.

Techniques: Real-time Polymerase Chain Reaction

Journal: Drug Design, Development and Therapy

Article Title: Cinobufagin Inhibits Invasion and Migration of Non-Small Cell Lung Cancer via Regulating Glucose Metabolism Reprogramming in Tumor-Associated Macrophages

doi: 10.2147/DDDT.S531190

Figure Lengend Snippet: Effect of CB on the Transcription Factor HIF-1α in M2 Macrophages. ( A ) Real-time quantitative PCR analysis of HIF-1α mRNA levels; ( B and C ) Western blot analysis of HIF-1α protein expression; ( D – F ) Western blot analysis of HIF-1α and Hydroxy-HIF-1α protein expression; ( G ) Immunoprecipitation analysis of HIF-1α ubiquitination levels. n=3; * P <0.05, ** P <0.01 compared to M0; # P <0.05 compared to M2; $ P <0.05 compared to M2+DMOG.

Article Snippet: Primary antibodies anti-HIF-1α (1:200; Cat# 66730-1-Ig; Proteintech) or anti-PD-L1 (1:100; Cat# 17952-1-AP; Proteintech) were added followed by overnight incubation at 4°C.

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Immunoprecipitation, Ubiquitin Proteomics

Journal: Drug Design, Development and Therapy

Article Title: Cinobufagin Inhibits Invasion and Migration of Non-Small Cell Lung Cancer via Regulating Glucose Metabolism Reprogramming in Tumor-Associated Macrophages

doi: 10.2147/DDDT.S531190

Figure Lengend Snippet: Effect of CB on the Transcription Factor HIF-1α in M2 Macrophages. ( A and B ) Immunofluorescence analysis of HIF-1α nuclear translocation (400×); ( C and D ) Fluorescence images of macrophages after uptake of 2-NBDG (400×); ( E ) Colorimetric measurement of lactate concentration; ( F – J ) Western blot analysis of protein expression levels for GLUT1, PKM2, LDHA, and MCT1. n=3; * P <0.05 compared to M0; # P <0.05 compared to M2; $ P <0.05 compared to M2+DMOG.

Article Snippet: Primary antibodies anti-HIF-1α (1:200; Cat# 66730-1-Ig; Proteintech) or anti-PD-L1 (1:100; Cat# 17952-1-AP; Proteintech) were added followed by overnight incubation at 4°C.

Techniques: Immunofluorescence, Translocation Assay, Fluorescence, Concentration Assay, Western Blot, Expressing

Journal: Drug Design, Development and Therapy

Article Title: Cinobufagin Inhibits Invasion and Migration of Non-Small Cell Lung Cancer via Regulating Glucose Metabolism Reprogramming in Tumor-Associated Macrophages

doi: 10.2147/DDDT.S531190

Figure Lengend Snippet: CB Targets HIF-1α to Regulate M2 Macrophage Polarization. ( A – C ) Real-time quantitative PCR analysis of mRNA levels for CD206, Arg-1, and IL-10; ( D and E ) ELISA measurement of TGF-β and IL-10 secretion levels. n=3; * P <0.05 compared to M0; # P <0.05 compared to M2; $ P <0.01 compared to M2+DMOG.

Article Snippet: Primary antibodies anti-HIF-1α (1:200; Cat# 66730-1-Ig; Proteintech) or anti-PD-L1 (1:100; Cat# 17952-1-AP; Proteintech) were added followed by overnight incubation at 4°C.

Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Drug Design, Development and Therapy

Article Title: Cinobufagin Inhibits Invasion and Migration of Non-Small Cell Lung Cancer via Regulating Glucose Metabolism Reprogramming in Tumor-Associated Macrophages

doi: 10.2147/DDDT.S531190

Figure Lengend Snippet: CB Targets HIF-1α to Regulate PD-L1 Expression in M2 Macrophages. ( A and B ) Western blot analysis of PD-L1 protein expression; ( C and D ) Immunofluorescence analysis of PD-L1 fluorescence intensity (400×); ( E and F ) Agarose gel electrophoresis of Input, IgG, and ChIP results. n=3; * P <0.05 compared to M0; # P <0.05, ## P <0.01 compared to M2; $ P <0.05 compared to M2+DMOG.

Article Snippet: Primary antibodies anti-HIF-1α (1:200; Cat# 66730-1-Ig; Proteintech) or anti-PD-L1 (1:100; Cat# 17952-1-AP; Proteintech) were added followed by overnight incubation at 4°C.

Techniques: Expressing, Western Blot, Immunofluorescence, Fluorescence, Agarose Gel Electrophoresis

Journal: Frontiers in Chemistry

Article Title: HIF1A acts as target of XiHuang Pill in the treatment of papillary thyroid cancer by regulating dedifferentiation

doi: 10.3389/fchem.2025.1607067

Figure Lengend Snippet: Analysis of molecular docking. The optimal binding conformation between (A) MOL001002 and AKT1, (B) MOL001002 and CCND1, (C) MOL001002 and ESR1, (D) MOL001027 and ESR1, (E) MOL001027 and HIF1A, (F) MOL001046 and BCL2, (G) MOL001046 and ESR1, (H) MOL001063 and CCND1, (I) MOL001063 and ESR1, (J) MOL001063 and HIF1A, (K) MOL001069 and BCL2, (L) MOL001069 and ESR1.

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 h at room temperature, followed by incubation overnight at 4°C with primary antibodies against HIF1A (#ER 1802-41, Huabio, Hangzhou, China), CD97 (#HA722563, Huabio), NIS (#24324-1-AP, Proteintech, IL, United States), TG (#21714-1-AP, Proteintech), TPO (#ab109383, Abcam, United Kingdom), and GAPDH (#10494-1-AP, Proteintech).

Techniques: Binding Assay

Journal: Frontiers in Chemistry

Article Title: HIF1A acts as target of XiHuang Pill in the treatment of papillary thyroid cancer by regulating dedifferentiation

doi: 10.3389/fchem.2025.1607067

Figure Lengend Snippet: The association of HIF1A with the immune microenvironment in PTC. (A) The infiltration abundance differences of immune cells between HIF1A high and low expression groups. (B) The correlation between HIF1A expression and infiltration abundance of activated NK cells. (C) The correlation between TDS and infiltration abundance of activated NK cells.

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 h at room temperature, followed by incubation overnight at 4°C with primary antibodies against HIF1A (#ER 1802-41, Huabio, Hangzhou, China), CD97 (#HA722563, Huabio), NIS (#24324-1-AP, Proteintech, IL, United States), TG (#21714-1-AP, Proteintech), TPO (#ab109383, Abcam, United Kingdom), and GAPDH (#10494-1-AP, Proteintech).

Techniques: Expressing

Journal: Frontiers in Chemistry

Article Title: HIF1A acts as target of XiHuang Pill in the treatment of papillary thyroid cancer by regulating dedifferentiation

doi: 10.3389/fchem.2025.1607067

Figure Lengend Snippet: HIF1A inhibits differentiation of PTC cells. (A) mRNA expression of HIF1A, CD97, NIS, TG, and TPO. (B) Protein expression of HIF1A, CD97, NIS, TG, and TPO. **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 h at room temperature, followed by incubation overnight at 4°C with primary antibodies against HIF1A (#ER 1802-41, Huabio, Hangzhou, China), CD97 (#HA722563, Huabio), NIS (#24324-1-AP, Proteintech, IL, United States), TG (#21714-1-AP, Proteintech), TPO (#ab109383, Abcam, United Kingdom), and GAPDH (#10494-1-AP, Proteintech).

Techniques: Expressing

Journal: Frontiers in Chemistry

Article Title: HIF1A acts as target of XiHuang Pill in the treatment of papillary thyroid cancer by regulating dedifferentiation

doi: 10.3389/fchem.2025.1607067

Figure Lengend Snippet: XHP inhibits proliferation, migration, and invasion while promoting differentiation of TPC-1 cells. (A) Cell viability was detected using CCK-8. (B) Cell proliferation was determined using colony formation analysis. (C) Cell migration was determined using wound healing analysis; scale bar = 500 μm. (D) Cell invasion was determined using Transwell analysis; scale bar = 200 μm. (E) mRNA expression of HIF1A, CD97, NIS, TG, and TPO. (F) Protein expression of HIF1A, CD97, NIS, TG, and TPO. TPC-1 cells were treated with 10 μmol/L (XHP-L), 50 μmol/L (XHP-M), or 100 μmol/L (XHP-H) of XHP. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 h at room temperature, followed by incubation overnight at 4°C with primary antibodies against HIF1A (#ER 1802-41, Huabio, Hangzhou, China), CD97 (#HA722563, Huabio), NIS (#24324-1-AP, Proteintech, IL, United States), TG (#21714-1-AP, Proteintech), TPO (#ab109383, Abcam, United Kingdom), and GAPDH (#10494-1-AP, Proteintech).

Techniques: Migration, CCK-8 Assay, Expressing

Journal: Frontiers in Chemistry

Article Title: HIF1A acts as target of XiHuang Pill in the treatment of papillary thyroid cancer by regulating dedifferentiation

doi: 10.3389/fchem.2025.1607067

Figure Lengend Snippet: XHP inhibits TPC-1 cell malignant features and promotes differentiation by downregulating HIF1A expression. (A) Expression of HIF1A was detected using qRT-PCR. (B) Cell proliferation was determined using colony formation analysis. (C) Cell migration was determined using wound healing analysis; scale bar = 500 μm. (D) Cell invasion was determined using Transwell analysis; scale bar = 200 μm. (E) mRNA expression of CD97, NIS, TG, and TPO. (F) Protein expression of CD97, NIS, TG, and TPO. TPC-1 cells were treated with 100 μmol/L XHP and transfected with oe-HIF1A. *P < 0.05, ***P < 0.001.

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 h at room temperature, followed by incubation overnight at 4°C with primary antibodies against HIF1A (#ER 1802-41, Huabio, Hangzhou, China), CD97 (#HA722563, Huabio), NIS (#24324-1-AP, Proteintech, IL, United States), TG (#21714-1-AP, Proteintech), TPO (#ab109383, Abcam, United Kingdom), and GAPDH (#10494-1-AP, Proteintech).

Techniques: Expressing, Quantitative RT-PCR, Migration, Transfection