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ay9944 (MedChemExpress)

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A Western blot analysis of SLC7A11, GPX4, and ACSL4 in SW480 (shNC and sh AURKA ) cells. B , C RNA-seq with KEGG enrichment analysis in SW480 (shNC and sh AURKA ) cells showing upregulated pathways in sh AURKA group ( B ), and a heatmap depicting DEGs in sterol/cholesterol biosynthesis pathways ( C ). D Schematic of the cholesterol biosynthesis pathway and its regulatory enzymes. E qPCR quantification of cholesterol biosynthesis enzyme mRNA levels in CRC (shNC and sh AURKA ) cells. F , G Western blot analysis of DHCR7 protein expression in SW480 and RKO cells ( F ), and SW480 cells following siRNA knockdown ( G ). H IC 50 analysis of RSL3 in SW480 (shNC and sh AURKA ) cells treated with RSL3 (48 h) following DHCR7 silencing. I Representative FACS images and quantified values of lipid ROS levels in SW480 (shNC and sh AURKA ) cells treated with RSL3 (5 μM, 24 h) following DHCR7 silencing. J LC-MS/MS quantification of 7-DHC levels in CRC (shNC and sh AURKA ) cells. K Quantification of RSL3 IC 50 values in SW480 (shNC and sh AURKA ) cells after pretreatment with 7-DHC (25 μM), cholesterol (25 μM) for 24 h. L Diagram of cholesterol synthesis showing DHCR7 converting 7-DHC to cholesterol and its inhibition by <t>AY9944.</t> M Intracellular levels of 7-DHC in SW480 (shNC and sh AURKA ) cells treated with AY9944 (100 nM) for 24 h. N Quantification of RSL3 IC 50 values in SW480 (shNC and sh AURKA ) cells after pretreatment with AY9944 (100 nM) for 24 h. O, P Quantification of intracellular lipid ROS ( O ) and MDA ( P ) levels in SW480 (shNC and sh AURKA ) cells treated with AY9944 (100 nM) for 24 h. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test [( E ), ( I ), ( J ), ( M , O , P )]. * p < 0.05, ** p < 0.01, *** p < 0.001.

Ay9944, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ay9944/AY+9944/pmc13039981-41-0-10
Average 94 stars, based on 14 article reviews

ay9944 - by Bioz Stars, 2026-10

94/100 stars

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1) Product Images from "AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer"

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

Journal: Cell Death & Disease

doi: 10.1038/s41419-026-08549-9

Figure Legend Snippet: A Western blot analysis of SLC7A11, GPX4, and ACSL4 in SW480 (shNC and sh AURKA ) cells. B , C RNA-seq with KEGG enrichment analysis in SW480 (shNC and sh AURKA ) cells showing upregulated pathways in sh AURKA group ( B ), and a heatmap depicting DEGs in sterol/cholesterol biosynthesis pathways ( C ). D Schematic of the cholesterol biosynthesis pathway and its regulatory enzymes. E qPCR quantification of cholesterol biosynthesis enzyme mRNA levels in CRC (shNC and sh AURKA ) cells. F , G Western blot analysis of DHCR7 protein expression in SW480 and RKO cells ( F ), and SW480 cells following siRNA knockdown ( G ). H IC 50 analysis of RSL3 in SW480 (shNC and sh AURKA ) cells treated with RSL3 (48 h) following DHCR7 silencing. I Representative FACS images and quantified values of lipid ROS levels in SW480 (shNC and sh AURKA ) cells treated with RSL3 (5 μM, 24 h) following DHCR7 silencing. J LC-MS/MS quantification of 7-DHC levels in CRC (shNC and sh AURKA ) cells. K Quantification of RSL3 IC 50 values in SW480 (shNC and sh AURKA ) cells after pretreatment with 7-DHC (25 μM), cholesterol (25 μM) for 24 h. L Diagram of cholesterol synthesis showing DHCR7 converting 7-DHC to cholesterol and its inhibition by AY9944. M Intracellular levels of 7-DHC in SW480 (shNC and sh AURKA ) cells treated with AY9944 (100 nM) for 24 h. N Quantification of RSL3 IC 50 values in SW480 (shNC and sh AURKA ) cells after pretreatment with AY9944 (100 nM) for 24 h. O, P Quantification of intracellular lipid ROS ( O ) and MDA ( P ) levels in SW480 (shNC and sh AURKA ) cells treated with AY9944 (100 nM) for 24 h. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test [( E ), ( I ), ( J ), ( M , O , P )]. * p < 0.05, ** p < 0.01, *** p < 0.001.

Techniques Used: Western Blot, RNA Sequencing, Expressing, Knockdown, Liquid Chromatography with Mass Spectroscopy, Inhibition

Software:

Article Title: Inhibiting cholesterol synthesis halts rhabdomyosarcoma growth via ER stress and cell cycle arrest.
Article Snippet: To explore the impact of HMGCR and DHCR7 chemical inhibition on cell proliferation, 2000–4000 cells were plated in 96- well plates (Corning) with five to six wells per condition. .. After 24 h, cells were exposed to varying concentrations of Lovastatin (# HYN0504, MCE) or AY9944 (DHCR7 inhibitor) (# HY-107420, MCE), and images were captured every 2 h to monitor growth dynamics using IncuCyte software. ..

Article Title: Targeting de novo cholesterol synthesis in rhabdomyosarcoma induces cell cycle arrest and triggers apoptosis through ER stress-mediated pathways
Article Snippet: To explore the impact of HMGCR and DHCR7 chemical inhibition on cell proliferation, 1000 - 2000 RD and KLHEL1 cells were plated in 96-well plates (Corning) with five replicates per condition. .. After 24 hours, cells were exposed to varying concentrations of Lovastatin (# HY- N0504, MCE) or AY9944 (DHCR7 inhibitor) (# HY-107420, MCE), and images were captured every 2 hours to monitor growth dynamics using IncuCyte software. ..

Article Title: Inhibiting cholesterol synthesis halts rhabdomyosarcoma growth via ER stress and cell cycle arrest
Article Snippet: To explore the impact of HMGCR and DHCR7 chemical inhibition on cell proliferation, 2000–4000 cells were plated in 96-well plates (Corning) with five to six wells per condition. .. After 24 h, cells were exposed to varying concentrations of Lovastatin (# HY-N0504, MCE) or AY9944 (DHCR7 inhibitor) (# HY-107420, MCE), and images were captured every 2 h to monitor growth dynamics using IncuCyte software. ..

other:

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer
Article Snippet: AY9944 (HY-107420), 7-Dehydrocholesterol (HY-113279), and Cholesterol (HY-N0322) were obtained from MedChemExpress (Shanghai, China).

Article Title: Inhibiting cholesterol synthesis halts rhabdomyosarcoma growth via ER stress and cell cycle arrest
Article Snippet: AY9944 , MedChemExpress , HY-107420.

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer.
Article Snippet: Reagents RSL3 (S8155), Erastin (S7242), FIN56 (S8254), 5-Fluorouracil (S1209), Oxaliplatin (S1224), Alisertib (S1133), AICAR (S1802) and Bafilomycin A1 (S1413) were obtained from Selleck (Houston, USA).

Cell Culture:

Article Title: Spatial proteomics of ovarian cancer precursors delineates early disease changes and drug targets.
Article Snippet: .. For OAW-42, OVCAR-8, and ES-2, we plated 4000 cells/well, and for EFO-21, we plated 6000 cells/well and we cultured the cells in full growth medium for 24 h. The medium was then removed and replaced with new full medium containing different concentrations of AY9944 (MedChemExpress, #HY-107420), carboplatin (Merck, #C2538) or combinations of the two drugs. .. Clinical data were obtained from the Tumor Bank Ovarian Cancer Network (www.toc-network.de) or the Charité Comprehensive Cancer Center (https://cccc.charite.de).

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Journal: Cell Death & Disease

Article Title: AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer

doi: 10.1038/s41419-026-08549-9

Figure Lengend Snippet: A Western blot analysis of SLC7A11, GPX4, and ACSL4 in SW480 (shNC and sh AURKA ) cells. B , C RNA-seq with KEGG enrichment analysis in SW480 (shNC and sh AURKA ) cells showing upregulated pathways in sh AURKA group ( B ), and a heatmap depicting DEGs in sterol/cholesterol biosynthesis pathways ( C ). D Schematic of the cholesterol biosynthesis pathway and its regulatory enzymes. E qPCR quantification of cholesterol biosynthesis enzyme mRNA levels in CRC (shNC and sh AURKA ) cells. F , G Western blot analysis of DHCR7 protein expression in SW480 and RKO cells ( F ), and SW480 cells following siRNA knockdown ( G ). H IC 50 analysis of RSL3 in SW480 (shNC and sh AURKA ) cells treated with RSL3 (48 h) following DHCR7 silencing. I Representative FACS images and quantified values of lipid ROS levels in SW480 (shNC and sh AURKA ) cells treated with RSL3 (5 μM, 24 h) following DHCR7 silencing. J LC-MS/MS quantification of 7-DHC levels in CRC (shNC and sh AURKA ) cells. K Quantification of RSL3 IC 50 values in SW480 (shNC and sh AURKA ) cells after pretreatment with 7-DHC (25 μM), cholesterol (25 μM) for 24 h. L Diagram of cholesterol synthesis showing DHCR7 converting 7-DHC to cholesterol and its inhibition by AY9944. M Intracellular levels of 7-DHC in SW480 (shNC and sh AURKA ) cells treated with AY9944 (100 nM) for 24 h. N Quantification of RSL3 IC 50 values in SW480 (shNC and sh AURKA ) cells after pretreatment with AY9944 (100 nM) for 24 h. O, P Quantification of intracellular lipid ROS ( O ) and MDA ( P ) levels in SW480 (shNC and sh AURKA ) cells treated with AY9944 (100 nM) for 24 h. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using unpaired Student’s t test [( E ), ( I ), ( J ), ( M , O , P )]. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: AY9944 (HY-107420), 7-Dehydrocholesterol (HY-113279), and Cholesterol (HY-N0322) were obtained from MedChemExpress (Shanghai, China).

Techniques: Western Blot, RNA Sequencing, Expressing, Knockdown, Liquid Chromatography with Mass Spectroscopy, Inhibition

Pharmacological inhibition of DHCR7 inhibits MetALD and HCC in DEN/HFD + EtOH-fed WT mice. DEN/HFD + EtOH-fed WT mice (n ≥10 per group) were therapeutically administered ±AY9944 (10 mg/kg, i.p., three times a week) for the last 9 weeks of feeding. (A) Levels of total serum, hepatic cholesterol, serum bile acids, and vitamin D 3 were measured. (B) Expression of Dhcr7 and Dhcr2 4 mRNA was measured using qRT-PCR. (C) Gross liver images of WT and Dhcr7 +/– mice. Tumor number, diameter, and burden were calculated. (D) Livers were stained for HCC markers AFP, YAP, and phospho-STAT3. (E) Livers were stained with H&E, Sirius red or immunostained for Desmin, αSMA, and F4/80. Area of positive staining was calculated as a percentage. (F) Expression of fibrogenic and inflammatory genes was measured using qRT-PCR. Data are mean ± SD; ∗ p <0.05, ∗∗ p <0.01, ∗∗∗ p <0.001, two-tailed Student’s t test. DEN, diethylnitrosamine; DHCR7, 7-dehydrocholesterol reductase; EtOH, ethanol; HCC, hepatocellular carcinoma ; HFD, high-fat diet; MetALD, metabolic dysfunction and alcohol-associated liver disease; qRT-PCR, quantitative real-time PCR; WT, wild-type.

Journal: JHEP Reports

Article Title: DHCR7 inhibition ameliorates MetALD and HCC in mice and human 3D liver spheroids

doi: 10.1016/j.jhepr.2025.101415

Figure Lengend Snippet: Pharmacological inhibition of DHCR7 inhibits MetALD and HCC in DEN/HFD + EtOH-fed WT mice. DEN/HFD + EtOH-fed WT mice (n ≥10 per group) were therapeutically administered ±AY9944 (10 mg/kg, i.p., three times a week) for the last 9 weeks of feeding. (A) Levels of total serum, hepatic cholesterol, serum bile acids, and vitamin D 3 were measured. (B) Expression of Dhcr7 and Dhcr2 4 mRNA was measured using qRT-PCR. (C) Gross liver images of WT and Dhcr7 +/– mice. Tumor number, diameter, and burden were calculated. (D) Livers were stained for HCC markers AFP, YAP, and phospho-STAT3. (E) Livers were stained with H&E, Sirius red or immunostained for Desmin, αSMA, and F4/80. Area of positive staining was calculated as a percentage. (F) Expression of fibrogenic and inflammatory genes was measured using qRT-PCR. Data are mean ± SD; ∗ p <0.05, ∗∗ p <0.01, ∗∗∗ p <0.001, two-tailed Student’s t test. DEN, diethylnitrosamine; DHCR7, 7-dehydrocholesterol reductase; EtOH, ethanol; HCC, hepatocellular carcinoma ; HFD, high-fat diet; MetALD, metabolic dysfunction and alcohol-associated liver disease; qRT-PCR, quantitative real-time PCR; WT, wild-type.

Article Snippet: AY9944 (in PBS; Cayman) was used for the inhibition of DHCR7 activity in vitro .

Techniques: Inhibition, Expressing, Quantitative RT-PCR, Staining, Two Tailed Test, Real-time Polymerase Chain Reaction

Administration of AY9944 inhibitor suppresses tumor growth in MetALD WT mice and MetALD-HCC human liver spheroids. (A) WT mice were fed HFD + EtOH for 2 weeks, adoptively transplanted with GFP + Hepa1-6 cells (1 × 10 6 ) via intrasplenic injection, followed by immediate splenectomy, and fed with HFD + EtOH for an additional 2 weeks. The AY9944 inhibitor was therapeutically administered (10 mg/kg, i.p., three times) during the last week of feeding. (B) Gross liver images of chimeric WT mice ± AY9944. Tumor number, diameter, and burden were calculated. (C) Representative liver images are shown, and GFP + tumors were visualized using fluorescent microscopy. (D) The number of tumors per lobe was calculated. (E) GFP signal was calculated in the left lobe as mean fluorescent intensity. (F) Liver function was assessed by measuring serum levels of ALT. (G) Levels of total serum and hepatic cholesterol were measured. (H) Expression of inflammatory genes was measured by qRT-PCR. (I) MetALD-HCC human liver spheroids were generated using HepG2 cells (3 × 10 5 cells transfected ± DHCR7 -targeting dsiRNA), NPCs, and HSCs and cultured in a MetALD cocktail for 7 days. (J) dsiRNAs with three different hairpins were tested for DHCR7 knockdown. (K) MetALD-HCC human liver spheroids (bright field images; scale bar = 200 μm). Relative volume of the HCC human liver spheroids was calculated as fold change compared with MetALD-HCC human liver spheroids containing HepG2 cells transfected with dsiRNA negative control. (L) Expression of HCC and proliferation markers was measured by qRT-PCR. (M) Expression of AFP was evaluated in MetALD-HCC human liver spheroids by Western blotting. Data are mean ± SD, ∗ p <0.05, ∗∗ p <0.01, ∗∗∗ p <0.001, two-tailed Student’s t test. ALT, alanine aminotransferase; DEN, diethylnitrosamine; DHCR7, 7-dehydrocholesterol reductase; EtOH, ethanol; HCC, hepatocellular carcinoma; HFD, high-fat diet; HSC, hepatic stellate cell; MetALD, metabolic dysfunction and alcohol-associated liver disease; NPC, non-parenchymal cell; qRT-PCR, quantitative real-time PCR; WT, wild-type.

Journal: JHEP Reports

Article Title: DHCR7 inhibition ameliorates MetALD and HCC in mice and human 3D liver spheroids

doi: 10.1016/j.jhepr.2025.101415

Figure Lengend Snippet: Administration of AY9944 inhibitor suppresses tumor growth in MetALD WT mice and MetALD-HCC human liver spheroids. (A) WT mice were fed HFD + EtOH for 2 weeks, adoptively transplanted with GFP + Hepa1-6 cells (1 × 10 6 ) via intrasplenic injection, followed by immediate splenectomy, and fed with HFD + EtOH for an additional 2 weeks. The AY9944 inhibitor was therapeutically administered (10 mg/kg, i.p., three times) during the last week of feeding. (B) Gross liver images of chimeric WT mice ± AY9944. Tumor number, diameter, and burden were calculated. (C) Representative liver images are shown, and GFP + tumors were visualized using fluorescent microscopy. (D) The number of tumors per lobe was calculated. (E) GFP signal was calculated in the left lobe as mean fluorescent intensity. (F) Liver function was assessed by measuring serum levels of ALT. (G) Levels of total serum and hepatic cholesterol were measured. (H) Expression of inflammatory genes was measured by qRT-PCR. (I) MetALD-HCC human liver spheroids were generated using HepG2 cells (3 × 10 5 cells transfected ± DHCR7 -targeting dsiRNA), NPCs, and HSCs and cultured in a MetALD cocktail for 7 days. (J) dsiRNAs with three different hairpins were tested for DHCR7 knockdown. (K) MetALD-HCC human liver spheroids (bright field images; scale bar = 200 μm). Relative volume of the HCC human liver spheroids was calculated as fold change compared with MetALD-HCC human liver spheroids containing HepG2 cells transfected with dsiRNA negative control. (L) Expression of HCC and proliferation markers was measured by qRT-PCR. (M) Expression of AFP was evaluated in MetALD-HCC human liver spheroids by Western blotting. Data are mean ± SD, ∗ p <0.05, ∗∗ p <0.01, ∗∗∗ p <0.001, two-tailed Student’s t test. ALT, alanine aminotransferase; DEN, diethylnitrosamine; DHCR7, 7-dehydrocholesterol reductase; EtOH, ethanol; HCC, hepatocellular carcinoma; HFD, high-fat diet; HSC, hepatic stellate cell; MetALD, metabolic dysfunction and alcohol-associated liver disease; NPC, non-parenchymal cell; qRT-PCR, quantitative real-time PCR; WT, wild-type.

Article Snippet: AY9944 (in PBS; Cayman) was used for the inhibition of DHCR7 activity in vitro .

Techniques: Injection, Microscopy, Expressing, Quantitative RT-PCR, Generated, Transfection, Cell Culture, Knockdown, Negative Control, Western Blot, Two Tailed Test, Real-time Polymerase Chain Reaction

Inhibition of DHCR7 suppresses the development of MASH and MetALD in human liver spheroids. (A) MASH- or MetALD-induced human liver spheroids were generated using primary steatotic hepatocytes, NPCs, and HSCs, and cultured in MASH or MetALD cocktails for 9 days ± AY9944 (administered during the last 4 days of culture). (B) Micrographs represent human liver spheroids stained with BODIPY and DAPI (scale bar = 250 μm). mRNA expression of (C) fatty acid-modifying enzymes, (D) CYP2E1, or (E) fibrogenic markers in MASH or MetALD human liver spheroids ( vs . control human liver spheroids). (F) Protein expression of fibrogenic markers in MASH or MetALD human liver spheroids detected by Western blotting. (G) Outline of AY9944 treatment of human liver spheroids with MetALD. (H) MetALD human liver spheroids (representative bright field images; scale bar = 200 μm). Expression of fibrogenic genes in AY9944-treated MetALD spheroids was measured by (I) qRT-PCR or (J) Western blotting. (K) Outline of AY9944 treatment of human liver spheroids with MASH. (L) MASH human liver spheroids (representative bright field images; scale bar = 200 μm). Expression of fibrogenic genes in AY9944-treated MASH spheroids was measured by (M) qRT-PCR or (N) Western blotting. DHCR7, 7-dehydrocholesterol reductase; HSC, hepatic stellate cell; MASH, metabolic dysfunction-associated steatohepatitis; MetALD, metabolic dysfunction and alcohol-associated liver disease; NPC, non-parenchymal cell; qRT-PCR, quantitative real-time PCR.

Journal: JHEP Reports

Article Title: DHCR7 inhibition ameliorates MetALD and HCC in mice and human 3D liver spheroids

doi: 10.1016/j.jhepr.2025.101415

Figure Lengend Snippet: Inhibition of DHCR7 suppresses the development of MASH and MetALD in human liver spheroids. (A) MASH- or MetALD-induced human liver spheroids were generated using primary steatotic hepatocytes, NPCs, and HSCs, and cultured in MASH or MetALD cocktails for 9 days ± AY9944 (administered during the last 4 days of culture). (B) Micrographs represent human liver spheroids stained with BODIPY and DAPI (scale bar = 250 μm). mRNA expression of (C) fatty acid-modifying enzymes, (D) CYP2E1, or (E) fibrogenic markers in MASH or MetALD human liver spheroids ( vs . control human liver spheroids). (F) Protein expression of fibrogenic markers in MASH or MetALD human liver spheroids detected by Western blotting. (G) Outline of AY9944 treatment of human liver spheroids with MetALD. (H) MetALD human liver spheroids (representative bright field images; scale bar = 200 μm). Expression of fibrogenic genes in AY9944-treated MetALD spheroids was measured by (I) qRT-PCR or (J) Western blotting. (K) Outline of AY9944 treatment of human liver spheroids with MASH. (L) MASH human liver spheroids (representative bright field images; scale bar = 200 μm). Expression of fibrogenic genes in AY9944-treated MASH spheroids was measured by (M) qRT-PCR or (N) Western blotting. DHCR7, 7-dehydrocholesterol reductase; HSC, hepatic stellate cell; MASH, metabolic dysfunction-associated steatohepatitis; MetALD, metabolic dysfunction and alcohol-associated liver disease; NPC, non-parenchymal cell; qRT-PCR, quantitative real-time PCR.

Article Snippet: AY9944 (in PBS; Cayman) was used for the inhibition of DHCR7 activity in vitro .

Techniques: Inhibition, Generated, Cell Culture, Staining, Expressing, Control, Western Blot, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

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ay9944 (MedChemExpress)

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1) Product Images from "AURKA-mediated destabilization of SAPS3 drives ferroptosis evasion via 7-dehydrocholesterol biosynthesis in colorectal cancer"

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