primary antibodies against hif 1α  (Proteintech)

Journal: Frontiers in Pharmacology

Article Title: Total alkaloids of Leonurus alleviate allergic asthma and inflammation responses by inhibiting hypoxia-induced factor-1⍺-mediated mast cell activation

doi: 10.3389/fphar.2026.1783196

Figure Lengend Snippet: HIF-1α is involved in the mechanism underlying TAL’s anti-asthma effects. (A) Schematic diagram of the cross-species transcriptomics. KEGG enrichment analysis of DEGs between DMSO- and TAL-groups (rno). KEGG enrichment analysis (hsa) of intersecting genes derived from DEGs and severe asthma (SA)-associated modules identified by WGCNA. Transcriptomic data were obtained from the GSE76262 clinical dataset. Venn diagram showing intersecting KEGG pathways between rat and human datasets. PPI network based on intersecting genes. (B) Boxplot showing HIF-1α expression levels in healthy controls (HC) versus SA patients. (C) ROC curve analysis of HIF-1⍺ in the GSE76262 dataset, evaluating its diagnostic performance in severe asthma. Transcriptomic data were obtained from the GSE76262 clinical dataset.

Article Snippet: Primary antibodies against anti-HIF-1α (1:1000, Cell Signaling Technology, 36169S), anti-β-actin (1:1000, Cell Signaling Technology, 4967S), and anti-GAPDH (1:1000, 2118S) followed by HRP-conjugated secondary antibodies (1:2500).

Techniques: Transcriptomics, Derivative Assay, Expressing, Diagnostic Assay

Journal: Frontiers in Pharmacology

Article Title: Total alkaloids of Leonurus alleviate allergic asthma and inflammation responses by inhibiting hypoxia-induced factor-1⍺-mediated mast cell activation

doi: 10.3389/fphar.2026.1783196

Figure Lengend Snippet: TAL inhibits HIF-1α expression and mast cell activation. (A) HIF-1α protein expression in RBL-2H3 cells under CoCl 2 (100 μM) induced-hypoxic with or without TAL treatment (0.5–2 mg/mL). (B) HIF-1α protein levels in lung tissues. (C) Immunofluorescence images and quantification of mean fluorescence intensity of HIF-1α in RBL-2H3 cells under hypoxic with or without TAL treatment (2 mg/mL). (D) β-hex release in RBL-2H3 cells following DNP-HSA stimulation and TAL treatment (0.5–2 mg/mL). (E,F) Levels of IL-4 (E) and IL-13 (F) secretion in RBL-2H3 cells following DNP-HSA stimulation and TAL treatment. Data are presented as mean ± SEM. from three independent experiments. ## P < 0.01, ### P < 0.001 compared with the normal group. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the OVA or model group.

Article Snippet: Primary antibodies against anti-HIF-1α (1:1000, Cell Signaling Technology, 36169S), anti-β-actin (1:1000, Cell Signaling Technology, 4967S), and anti-GAPDH (1:1000, 2118S) followed by HRP-conjugated secondary antibodies (1:2500).

Techniques: Expressing, Activation Assay, Immunofluorescence, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Total alkaloids of Leonurus alleviate allergic asthma and inflammation responses by inhibiting hypoxia-induced factor-1⍺-mediated mast cell activation

doi: 10.3389/fphar.2026.1783196

Figure Lengend Snippet: HIF-1α promotes mast cell activation and attenuates TAL’s inhibitory effect. (A,B) Western blot (A) and RT-PCR (B) validation of the levels of HIF-1α in RBL-2H3 cells. (C) Western blot analysis of HIF-1α protein expression in EV and HIF-1α-overexpressing cells with or without TAL treatment. (D–F) Levels of β-hex (D) , IL-4 (E) , and IL-13 (F) in the indicated groups following DNP-HSA stimulation and TAL treatment. (G–I) Effect of the HIF-1α inhibitor YC-1 (20 µM) on β-hex release (G) , IL-4 (H) , and IL-13 (I) secretion in DNP-HSA-stimulated RBL-2H3 cells. Data are expressed as mean ± SEM from three independent experiments. ## P < 0.01, ### P < 0.001 compared with the control. * P < 0.05, ** P < 0.01, *** P < 0.001compared with the DNP-HSA simulated group or as indicated.

Article Snippet: Primary antibodies against anti-HIF-1α (1:1000, Cell Signaling Technology, 36169S), anti-β-actin (1:1000, Cell Signaling Technology, 4967S), and anti-GAPDH (1:1000, 2118S) followed by HRP-conjugated secondary antibodies (1:2500).

Techniques: Activation Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Expressing, Control

Journal: bioRxiv

Article Title: A tumor metabolism-angiogenesis-immune axis governs immunotherapy responses

doi: 10.64898/2026.02.23.707524

Figure Lengend Snippet: ( A ) Representative coronal and transaxial fused PET/CT rendering (PET in colormap, CT in greyscale) and tumor biodistribution of in vivo glucose uptake using [ 18 F]FDG in mice (n=4) bearing 4T1-KD and 4T1-Sc tumors in contralateral m.f.p. at day 11-14 of tumor growth to equalize tumor volumes. ( B ) Quantification and representative images of pixel intensity for glucose detected by MALDI imaging metabolomics in 4T1-KD and 4T1-Sc tumor tissue. ( C ) Metabolic analysis of EC isolated from Sc and KD tumors by Seahorse, showing extracellular acidification rates (ECAR) and oxygen consumption rates (OCR), and ( D ) Seahorse analysis of Sc- and KD-derived ECs treated with or without murine recombinant (mr)VEGF-A (10 ng/ml). Significance is represented with (*) for control Sc-ECs vs. control KD-ECs, and with (#) for control KD-ECs vs KD-ECs+mrVEGF-A ((n=8/group; Data are mean +/- SD from one representative out of 2-3 independent experiments). ( E-H ) IF quantification of % CD31 ECs (E), fractions of CD31 + vessels covered by PCs (F), vasculature complexity by mean CD31 + cell numbers in CD31 + vessel structures (G), % HIF-1a + cells (H) in 4T1-Sc and KD tumors (n=3-5). ( I, J ) Vessel leakiness by Evans blue assay in 4T1-Sc and 4T1-KD tumors on day 10 and 13 of growth respectively (I), and (J) in vivo HIF-1 reporter activity in day 7 vs. day 10 4T1-Sc and 4T1-KD tumors injected 3 days apart in contralateral m.f.p. of WT mice (n=17, 3 experiments combined). ( K ) Representative DCE MR images (showing contralateral 4T1-Sc and 4T1-KD tumors; enlarged dLNs; filled bladder, B; and rectum, R, with inserted rectal temperature probe) (left), and representative average normalized signal-time curves of contrast agent uptake (wash-in and wash-out) behavior of well-vascularized (Pattern 1, typically well-oxygenated), vascularized (Pattern 2, hypoxic) and non-vascularized (Pattern 3, includes necrotic) tumor areas (middle), and (right) related quantification of volume fractions of the indicated tumor perfusion patterns in 4T1-Sc and 4T1-KD tumor tissue (n=12, 2 separate cohorts combined; dotted white line indicates patterns that are significantly decreased in 4T1-KD vs. 4T1-Sc tumors). ( L,M ) Experiment schema with tumor volume (L), and (M) flow cytometry quantification of tumor infiltrating T cells upon adoptive transfer of tumor-specific T cells into RAG2 KO mice bearing contralateral orthotopic 4T1-Sc and 4T1-KD tumors. ( N ) Flow cytometry quantification of proportions of tumor converted T cells (Kaede red + ) recovered in dLN relative to the same converted T-cell subsets in tumors (TM) of Kaede mice implanted with B16-Sc or B16-KD (n=4/each). Data are mean +/- SE from one representative out of 2-3 independent experiments. 2-sided unpaired (A-H, N) or paired (I-M) t test. *, P<0.05; **,P<0.01; ***, P<0.001; ****P<0.0001.

Article Snippet: Primary antibodies against HIF-1α (0.5 μg/ml, Novus Bio, cat#NB100-479), NG2 (5 μg/ml, Millipore, cat#AB5320), and CD31 (0.08 μg/ml, Abcam, cat#ab182981) were incubated for 1 hour, followed by 8 minutes incubation with Leica Bond Polymer anti-rabbit HRP (included in the Polymer Refine Detection Kit (Leica, cat#DS9800).

Techniques: Positron Emission Tomography-Computed Tomography, In Vivo, Imaging, Isolation, Derivative Assay, Recombinant, Control, Evans Blue Assay, Activity Assay, Injection, Flow Cytometry, Adoptive Transfer Assay

Journal: iScience

Article Title: Young rat vascular endothelial cells promote neurological recovery of stroke aged rat via HIF-1α

doi: 10.1016/j.isci.2025.112552

Figure Lengend Snippet: The protein level, mRNA expression level, and DNA binding activity of HIF-1α in the ischemic penumbra at different time points There was no significant difference in protein (A and B) or mRNA levels (C) between young and aged rats after cerebral ischemic injury. The DNA binding activity of HIF-1α was significantly reduced in aged rats on days 1, 3, and 7 after MCAO compared with young rats (D). Data were presented as mean ± SD ( n = 5 rats per group), ∗ p < 0.05 (unpaired t-test).

Article Snippet: Then, the membrane was incubated with primary antibody against HIF-1α (1:1,000; Signalway Antibody, USA), NGF (1:1000; Santa Cruz, USA), or β-actin (1:1,000; Santa Cruz, USA) at 4°C.

Techniques: Expressing, Binding Assay, Activity Assay

Journal: iScience

Article Title: Young rat vascular endothelial cells promote neurological recovery of stroke aged rat via HIF-1α

doi: 10.1016/j.isci.2025.112552

Figure Lengend Snippet: The protein expression and DNA binding activity of HIF-1α in aged VECs and young VECs (A) The VECs were characterized by immunofluorescence technique. (B) The DNA binding activity of HIF-1α was reduced in aged VECs compared with young VECs. (C and D) No significant difference was observed in HIF-1α protein level between aged and young VECs, and the HIF-1α protein expression could be significantly inhibited by siRNA in the young VECs. Data were presented as mean ± SD ( n = 5 rats per group), ∗ p < 0.05 (unpaired t-test).

Article Snippet: Then, the membrane was incubated with primary antibody against HIF-1α (1:1,000; Signalway Antibody, USA), NGF (1:1000; Santa Cruz, USA), or β-actin (1:1,000; Santa Cruz, USA) at 4°C.

Techniques: Expressing, Binding Assay, Activity Assay, Immunofluorescence

Journal: iScience

Article Title: Young rat vascular endothelial cells promote neurological recovery of stroke aged rat via HIF-1α

doi: 10.1016/j.isci.2025.112552

Figure Lengend Snippet: Young VECs promoted neurologic function recovery of aged rats by lateral ventricle transplantation Aged rats with MCAO were treated with vehicle, aged VECs, young VECs or Young VEC with HIF-1α siRNA. (A) Compared with the other three groups, the MCAO + young VEC group showed significant greater neurological recovery. ∗ p < 0.05 vs. MCAO + young VEC (two-way repeated measures ANOVA). (B and C) At day 3 after MCAO, TTC staining was used to measure infarct volume. The infarct volumes of the MCAO + young VEC group were significantly decreased compared with that of the other four groups. n = 10 rats per group, ∗ p < 0.05 (two-way repeated measures ANOVA). Data were presented as mean ± SD.

Article Snippet: Then, the membrane was incubated with primary antibody against HIF-1α (1:1,000; Signalway Antibody, USA), NGF (1:1000; Santa Cruz, USA), or β-actin (1:1,000; Santa Cruz, USA) at 4°C.

Techniques: Transplantation Assay, Staining