vhl inhibitor vh298  (MedChemExpress)

Journal: EMBO Reports

Article Title: Conserved lipid metabolic reprogramming confers hypoxic and aging resilience

doi: 10.1038/s44319-025-00664-6

Figure Lengend Snippet: ( A ) Schematic illustrating fatty acid synthesis and desaturation pathways. ( B ) Representative confocal fluorescence images showing the expression of FASN-1::GFP under normal or hypoxic (0.1%) conditions for 24 h post L4 stages. Scale bars: 100 µm (low mag) or 10 µm (high mag). ( C ) Representative confocal fluorescence images showing high-resolution Z-stack views of fat-6p::fat-6::GFP under normal or hypoxic (0.1%) conditions for 24 h post L4 stages. Scale bars: 10 µm. ( D ) Representative confocal fluorescence images showing low and high-resolution Z-stack views of fat-6p::fat-6::GFP under normal or hypoxic (0.5%) conditions for 24 h post L4 stages. Scale bars: 10 µm. ( E ) Quantification of the percentage of animals with fat-6p::fat-6::GFP fluorescence intensities downregulated under normal or hypoxic (0.5%) conditions for 24 h post L4 stages. Data were presented as means ± S.D. P values calculated by unpaired two-tailed t-tests. *** P < 0.001 (normal, n = 30 animals, hypoxic, n = 37 animals). ( F ) Representative confocal fluorescence images showing the downregulation of dhs-3p::dhs-3::GFP -labeled lipid droplet abundance and number under normal or hypoxic (0.1%) conditions for 24 h post L4 stages. Scale bars: 10 µm. ( G ) Representative confocal fluorescence images showing the expression of HIF-1-dependent transcriptional reporter, nis470 (cysl-2p::mVenus) abundance under DMSO or VHL-1 inhibitor VH-298 (1 mg/ml) for 96 h post embryos stage. Scale bars: 10 µm. ( H ) Representative confocal fluorescence images showing low and high-resolution Z-stack views of fat-6p::fat-6::GFP under DMSO or VHL-1 inhibitor VH-298 (1 mg/ml) for 96 h post embryos stage. Scale bars: 10 µm. ( I ) Quantification of the percentage of animals with fat-6p::fat-6::GFP fluorescence intensities downregulated under DMSO or VHL-1 inhibitor VH-298 (1 mg/ml) for 96 h post embryos stage. Data were presented as means ± SEM. P values calculated by unpaired two-tailed t-tests. * P < 0.05 ( n = 30 animals per condition). ( J ) Representative confocal fluorescence images showing fat-6p::fat-6::GFP animals fed with control RNAi or RNAi against hif-1 or vhl-1 under normal or hypoxic conditions. Scale bars: 10 µm. ( K ) Representative confocal fluorescence images showing dhs-3p::dhs-3::GFP animals fed with control RNAi or RNAi against hif-1 or vhl-1 under normal conditions. Scale bars: 10 µm. ( L ) Representative confocal fluorescence images showing FASN-1::GFP animals fed with control RNAi or RNAi against vhl-1 under normal conditions. Scale bars: 10 µm. .

Article Snippet: VH-298 , MedChemExpress , Cat# HY-100947.

Techniques: Fluorescence, Expressing, Two Tailed Test, Labeling, Control

Journal: EMBO Reports

Article Title: Conserved lipid metabolic reprogramming confers hypoxic and aging resilience

doi: 10.1038/s44319-025-00664-6

Figure Lengend Snippet: ( A ) Representative confocal fluorescence images showing the expression of an integrated transgene fat-6p::fat-6::GFP under normal conditions at L1-L4 stages. Scale bars: 100 µm. ( B ) Representative Low and high magnification confocal fluorescence images showing the expression of fat-6p::fat-6::GFP under normal conditions L4 stages. Scale bars: 10 µm. ( C ) Representative confocal fluorescence images showing low and high-resolution Z-stack views of fat-6p::fat-6::GFP under normal or hypoxic (5%) conditions for 24 h post L4 stages. Scale bars: 10 µm. ( D ) Quantification of the percentage of animals with fat-6p::fat-6::GFP fluorescence intensities downregulated under normal or hypoxic (5%) conditions for 24 h post L4 stages. Data were presented as means ± S.D. P values calculated by unpaired two-tailed t-tests. n.s indicates nonsignificant ( n > 25 animals per condition). ( E ) Representative confocal fluorescence images showing fat-6p::fat-6::GFP fed with control RNAi or RNAi against hif-1 . Scale bars: 10 µm. ( F ) Representative confocal fluorescence images showing the downregulation of dhs-3p::dhs-3::GFP -labeled lipid droplet number and size under normal or hypoxic (0.5%) conditions for 24 h post-L4 stages. Scale bars: 10 µm. ( G ) Schematic diagram of HIF-1, VHL-1, and the drug VH-298. Representative confocal fluorescence images showing the upregulation of HIF-1 target reporter cysl-2p::Venus treat with DMSO or VHL-1 inhibitor VH-298 (1 mg/ml) for 48 h starting at embryos. Scale bars: 10 µm. ( H ) Representative confocal fluorescence images showing the upregulation of HIF-1 target reporter cysl-2p::Venus treat with DMSO or VHL-1 inhibitor VH-298 (10 mg/ml) for 48 h starting at embryos. Scale bars: 10 µm. ( I ) Representative confocal fluorescence images showing the upregulation of HIF-1 target reporter cysl-2p::Venus treat with DMSO or VHL-1 inhibitor VH-298 (10 mg/ml) for 96 h starting at embryos. Scale bars: 10 µm.

Article Snippet: VH-298 , MedChemExpress , Cat# HY-100947.

Techniques: Fluorescence, Expressing, Two Tailed Test, Control, Labeling

Journal: Nature Communications

Article Title: The urotensin II receptor triggers an early meningeal response and a delayed macrophage-dependent vasospasm after subarachnoid hemorrhage in male mice

doi: 10.1038/s41467-024-52654-2

Figure Lengend Snippet: a Immunolabeling of ERTR7 + (white) meningeal fibroblasts of leptomeninges showing UT (magenta) overexpression and DAPI (blue) in SAH compared with sham conditions. Quantification of UT intensity from D1 to D7 in ERTR7 + areas ( n = 5). Scale bar = 50 µm. b Immunolabeling of the dura mater showing UT (green) overexpression in SAH compared to sham conditions at D1 in ERTR7 + (magenta) meningeal fibroblasts bordering dural lectin + (gray) vessels ( n = 3/condition). Scale bar = 50 µm. c Immunolabeling of leptomeninges showing HIF-1α (cyan) overexpression at D1 in AKAP12 + (green) meningeal fibroblasts in SAH compared with sham conditions. Quantification of meningeal HIF-1α intensity in AKAP12 + areas ( n = 6). Scale bar = 50 µm. d Timeline of intracisternal injections of siRNA targeting HIF-1α gene expression one day before SAH surgery. e Immunolabeling of leptomeninges showing at D1 HIF-1α (green) and UT (magenta) expression in SAH compared to sham conditions in mice pretreated with siCTRL or siHIF-1α in laminin + (gray) meningeal fibroblasts. Quantification of HIF-1α (upper panel) and UT (lower panel) intensity in laminin + areas ( n = 4). Scale bar = 50 µm. f MCA delimited via lectin + (green) and DAPI (blue) staining at D7. MCA lumen area/wall thickness ratio in SAH compared to sham conditions in mice pretreated with siCTRL or siHIF-1α (lower panel). Quantification of the lumen area/wall thickness ratio ( n = 6). Scale bar = 50 µm. HIF-1α (magenta) expression in peri-MCA in SAH compared to sham UT +/+ and UT -/- mice (upper panel). Quantification of perivascular HIF-1α intensity in the MCA ( n = 6). Scale bar = 50 µm. g Timeline of intracisternal injection of aCSF or VH298 prior to intracisternal injection of aCSF or UII. h Immunolabeling of HIF-1α (red) or UT (cyan) within leptomeninges at D1 in sham- or VH298-pretreated mice and in the absence or presence of UII. Quantification of HIF-1α (upper panel) and UT (lower panel) intensity in meningeal cells. Scale bar = 50 µm. i MCA delineation with lectin (green) and DAPI (blue) staining at D7. Quantification of the lumen area/wall thickness ratio ( n = 3). Scale bar = 50 µm. a – i Values are expressed as the mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 ( a – g , i ) Two-sided two-way analysis of variance (ANOVA), Bonferroni’s correction). h two-sided two-way analysis of variance (ANOVA), Tukey’s correction). Source data are provided as a Source Data file. a , d , g created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license ( https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en ).

Article Snippet: In another experimental set, HIF-1α stabilization was performed by intracisternal injection of the Von Hippel‒Lindau (VHL) VH298 inhibitor (10 μl, 5.2 μg/ml) (Tocris, #6156) or aCSF volume as a control 2 h before intracisternal injection of murine UII (50 μl, 2 μg/ml) (Phoenix Pharmaceuticals, #071-08) or aCSF volume solution for two consecutive days (D-1, D0).

Techniques: Immunolabeling, Over Expression, Gene Expression, Expressing, Staining, Injection

Journal: Nature Communications

Article Title: The urotensin II receptor triggers an early meningeal response and a delayed macrophage-dependent vasospasm after subarachnoid hemorrhage in male mice

doi: 10.1038/s41467-024-52654-2

Figure Lengend Snippet: a Kinetics of F4/80 + (green) pial MΦ recruitment and UT (magenta) overexpression in leptomeninges from D1 to D7 in SAH compared to sham UT +/+ mice. Quantification of the number of F4/80 + MΦs (left panel, n = 5) and UT intensity in F4/80 + cells (right panel, n = 3) from D1 to D7. Scale bar = 50 µm. Values are expressed as the mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001 (normally distributed, comparison of two groups, unpaired two sided t -test). b FACS analysis and gating strategy (upper right quadrant F4/80 + UT + ) for bone marrow-derived (in vitro, see Material and Methods) F4/80 + MΦs (BMDMs) obtained from UT +/+ or UT -/- mice and treated or not treated with VH298 (24 h). Quantification of the percentage of UT-expressing cells among total F4/80 + BMDMs ( n = 3). Values are expressed as the mean ± SEM. *** P < 0.001 (two-sided two-way analysis of variance (ANOVA), Tuckey’s correction). c Immunolabeling of leptomeninges of UT (magenta) and F4/80 + (green) MΦs at D1 in aCSF- or VH298-pretreated mice and in the absence or presence of UII in the subarachnoid space ( n = 3). Scale bar = 50 µm. Values are expressed as the mean ± SEM. * P < 0.05 (two-sided two-way analysis of variance (ANOVA), Tuckey’s correction). d Experimental timeline of leptomeningeal and PVMΦ depletion by intracisternal injection of clodronate-liposomes (CLO-lip) before SAH with blood from mice depleted of peripheral MΦs prior to behavioral testing and brain analyses. Values are expressed as the mean ± SEM. * P < 0.05; *** P < 0.001 (two-sided two-way analysis of variance (ANOVA), Tuckey’s correction). e Immunolabeling of F4/80 + (magenta) MΦs around lectin + (white) leptomeningeal and perivascular vessels. Histograms of quantification of the total number of F4/80 + cells at D1 in sham and SAH conditions. f Immunolabeling of lectin + (green) and nuclei by DAPI (blue) in MCA at D7 in PBS-lip and CLO pretreated mice. Quantification of lumen area/wall thickness ( n = 6). Scale bar = 50 µm. g Exploration and locomotion in OFT. Sensorimotor functions in the BWT and preference index in the NORT ( n = 9/condition). Radar plots illustrating the relative effect of SAH in CLO pretreated mice on performance in OFT, BWT, and NOR test. Each item of the radar represents the mean normalized to CLO pretreated PBS mice. e – g Values are expressed as the mean ± SEM. ns=non-significant, * P < 0.05; ** P < 0.01; *** P < 0.001. (two-sided two-way analysis of variance (ANOVA), Bonferroni’s correction). Source data are provided as a Source Data file. b , d – f created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license ( https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en ).

Article Snippet: In another experimental set, HIF-1α stabilization was performed by intracisternal injection of the Von Hippel‒Lindau (VHL) VH298 inhibitor (10 μl, 5.2 μg/ml) (Tocris, #6156) or aCSF volume as a control 2 h before intracisternal injection of murine UII (50 μl, 2 μg/ml) (Phoenix Pharmaceuticals, #071-08) or aCSF volume solution for two consecutive days (D-1, D0).

Techniques: Over Expression, Comparison, Derivative Assay, In Vitro, Expressing, Immunolabeling, Injection, Liposomes