sphk1 activity assay Search Results


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  • 91
    Bethyl sphk1
    <t>SPHK1</t> is a novel target of metformin. (A) SPHK1 mRNA expression was measured by qRT-PCR in TYKnu (left) and CAOV3 (right) OvCa cells treated with metformin or phenformin at the indicated doses for 24h. Results expressed as relative quantity over GAPDH housekeeping gene (n=3) (B) SPHK1 protein expression was assessed by immunoblot in TYKnu (left) and CAOV3 (right) cells treated with metformin or phenformin at the indicated doses for 72h, (n=3). (C) Wound healing assay in OVCAR5 cells stably overexpressing SPHK1 (SPHK1-OE) (n=6). (D) MTT assay measuring proliferation in SPHK1 overexpressing and control transfected OVCAR5 cells (n=6). (E) Immunoblot of AKT activation (S473 phosphorylation) in SPHK1 overexpressing and control transfected OVCAR5 cells (n=3). (F) Colony formation assay showing number of colonies formed after 10 days in SPHK1 overexpressing and control transfected OVCAR5 cells (n=3). (G) SOX2 mRNA expression was measured by qRT-PCR in OVCAR5 cells overexpressing SPHK1 or control vector. Results expressed as relative quantity over GAPDH housekeeping gene (n=3). (H) SOX2 protein expression was assessed by immunoblot in OVCAR5 cells overexpressing SPHK1 or control vector. (I) Xenograft mouse model with OVCAR5 cells overexpressing of SPHK1 was compared to vector control OVCAR5 cells (Control n=9, SPHK1-OE n=10). X-axis is expressed as fold change in mean tumor weight (g) over control. IHC analysis of tumors from SPHK1 overexpressing group and control group measuring expression of Ki67 (J) and SOX2 (K) . Data represent mean value ± S.D. * p
    Sphk1, supplied by Bethyl, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sphk1/product/Bethyl
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sphk1 - by Bioz Stars, 2022-12
    91/100 stars
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    91
    Abnova sphk1
    MiR-124 directly targets <t>SphK1</t> and inhibits the migration and invasion of ovarian cancer cells. (A) Wound healing assay of SKOV3-ip and HO8910pm cells transfected with NC or miR-124 mimics. Representative pictures are shown at 0 and 24 h after the wound was made. (B) Transwell assay of SKOV3-ip and HO8910pm cells transfected with NC or miR-124 mimics. (C) Matrigel invasion assay of SKOV3-ip and HO8910pm cells transfected with miR-124 mimics or NC. (D) Expression of endogenous SphK1 in SKOV3-ip cells at 48 h and 72 h post transfection of miR-124 mimics or NC. β-actin was loaded as an internal control. (E) Diagram of the SphK1-3’-UTR with potential binding-sites for miR-124. (F) Relative luciferase activity of reporters including wild-type or mutant SphK1 3’-UTR co-transfected with NC or miR-124 mimics. ** and *** indicate significant difference at p
    Sphk1, supplied by Abnova, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sphk1/product/Abnova
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sphk1 - by Bioz Stars, 2022-12
    91/100 stars
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    92
    Cayman Chemical sphk1
    MiR-124 directly targets <t>SphK1</t> and inhibits the migration and invasion of ovarian cancer cells. (A) Wound healing assay of SKOV3-ip and HO8910pm cells transfected with NC or miR-124 mimics. Representative pictures are shown at 0 and 24 h after the wound was made. (B) Transwell assay of SKOV3-ip and HO8910pm cells transfected with NC or miR-124 mimics. (C) Matrigel invasion assay of SKOV3-ip and HO8910pm cells transfected with miR-124 mimics or NC. (D) Expression of endogenous SphK1 in SKOV3-ip cells at 48 h and 72 h post transfection of miR-124 mimics or NC. β-actin was loaded as an internal control. (E) Diagram of the SphK1-3’-UTR with potential binding-sites for miR-124. (F) Relative luciferase activity of reporters including wild-type or mutant SphK1 3’-UTR co-transfected with NC or miR-124 mimics. ** and *** indicate significant difference at p
    Sphk1, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sphk1/product/Cayman Chemical
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sphk1 - by Bioz Stars, 2022-12
    92/100 stars
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    95
    Echelon Biosciences sphk1 activity
    Impact of tamoxifen and LCL204 on AC and <t>Sphk1</t> expression and Sphk1 activity in AML cell lines. (A) Tamoxifen dose-response in KG-1 cells. KG-1 cells were seeded in 6-well plates, 5 × 10 6 /well, in media containing 5% FBS. After 60 min, cells were
    Sphk1 Activity, supplied by Echelon Biosciences, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sphk1 activity/product/Echelon Biosciences
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sphk1 activity - by Bioz Stars, 2022-12
    95/100 stars
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    Image Search Results


    SPHK1 is a novel target of metformin. (A) SPHK1 mRNA expression was measured by qRT-PCR in TYKnu (left) and CAOV3 (right) OvCa cells treated with metformin or phenformin at the indicated doses for 24h. Results expressed as relative quantity over GAPDH housekeeping gene (n=3) (B) SPHK1 protein expression was assessed by immunoblot in TYKnu (left) and CAOV3 (right) cells treated with metformin or phenformin at the indicated doses for 72h, (n=3). (C) Wound healing assay in OVCAR5 cells stably overexpressing SPHK1 (SPHK1-OE) (n=6). (D) MTT assay measuring proliferation in SPHK1 overexpressing and control transfected OVCAR5 cells (n=6). (E) Immunoblot of AKT activation (S473 phosphorylation) in SPHK1 overexpressing and control transfected OVCAR5 cells (n=3). (F) Colony formation assay showing number of colonies formed after 10 days in SPHK1 overexpressing and control transfected OVCAR5 cells (n=3). (G) SOX2 mRNA expression was measured by qRT-PCR in OVCAR5 cells overexpressing SPHK1 or control vector. Results expressed as relative quantity over GAPDH housekeeping gene (n=3). (H) SOX2 protein expression was assessed by immunoblot in OVCAR5 cells overexpressing SPHK1 or control vector. (I) Xenograft mouse model with OVCAR5 cells overexpressing of SPHK1 was compared to vector control OVCAR5 cells (Control n=9, SPHK1-OE n=10). X-axis is expressed as fold change in mean tumor weight (g) over control. IHC analysis of tumors from SPHK1 overexpressing group and control group measuring expression of Ki67 (J) and SOX2 (K) . Data represent mean value ± S.D. * p

    Journal: Molecular cancer research : MCR

    Article Title: SPHK1 is a novel target of metformin in ovarian cancer

    doi: 10.1158/1541-7786.MCR-18-0409

    Figure Lengend Snippet: SPHK1 is a novel target of metformin. (A) SPHK1 mRNA expression was measured by qRT-PCR in TYKnu (left) and CAOV3 (right) OvCa cells treated with metformin or phenformin at the indicated doses for 24h. Results expressed as relative quantity over GAPDH housekeeping gene (n=3) (B) SPHK1 protein expression was assessed by immunoblot in TYKnu (left) and CAOV3 (right) cells treated with metformin or phenformin at the indicated doses for 72h, (n=3). (C) Wound healing assay in OVCAR5 cells stably overexpressing SPHK1 (SPHK1-OE) (n=6). (D) MTT assay measuring proliferation in SPHK1 overexpressing and control transfected OVCAR5 cells (n=6). (E) Immunoblot of AKT activation (S473 phosphorylation) in SPHK1 overexpressing and control transfected OVCAR5 cells (n=3). (F) Colony formation assay showing number of colonies formed after 10 days in SPHK1 overexpressing and control transfected OVCAR5 cells (n=3). (G) SOX2 mRNA expression was measured by qRT-PCR in OVCAR5 cells overexpressing SPHK1 or control vector. Results expressed as relative quantity over GAPDH housekeeping gene (n=3). (H) SOX2 protein expression was assessed by immunoblot in OVCAR5 cells overexpressing SPHK1 or control vector. (I) Xenograft mouse model with OVCAR5 cells overexpressing of SPHK1 was compared to vector control OVCAR5 cells (Control n=9, SPHK1-OE n=10). X-axis is expressed as fold change in mean tumor weight (g) over control. IHC analysis of tumors from SPHK1 overexpressing group and control group measuring expression of Ki67 (J) and SOX2 (K) . Data represent mean value ± S.D. * p

    Article Snippet: Together, these data support the S1P pathway and SPHK1 in particular, as potential targets for OvCa treatment.

    Techniques: Expressing, Quantitative RT-PCR, Wound Healing Assay, Stable Transfection, MTT Assay, Transfection, Activation Assay, Colony Assay, Plasmid Preparation, Immunohistochemistry

    Metformin inhibits hypoxia-induced expression of SPHK1 in OvCa. (A) SPHK1 protein expression was assessed by immunoblot in OVCAR5, Kuramochi and TYKnu cells overexpressing constitutively stable HIF1α with P402A/P564A (left) or HIF2α with P405A/P531A (right) mutations (n=3). (B) SPHK1 protein expression was analyzed by immunoblot after metformin treatment (1mM, 72h) in OVCAR5 and Kuramochi cells overexpressing constitutively stable HIF1α with activating substitution P402A/P564A (left) or HIF2α with P405A/P531A (right) (n=3). (C) HIF1α, HIF2α and SPHK1 protein expression were assessed by immunoblot in response to hypoxia (1% O 2 , 5% CO 2 ) with metformin concurrent treatment (1mM, 48h) (n=3). (D) HIF1α and HIF2α protein expression and localization in OVCAR5 cells was measured by immunoblot following cellular fractionation. HIF expression was stabilized using CoCl 2 (100μM) and concurrently treated with metformin (1mM, 24h). Histone H3 and βTubulin were used to normalize nuclear and cytosolic localization, respectively (n=3). (E) HIF1α and HIF2α protein expression and localization in OVCAR5 cells was measured by immunoblot following cellular fractionation following transfection with HIF1α or HIF2α constitutively stable mutants and subsequent treatment with metformin (1mM, 24h). Histone H3 and βTubulin were used to confirm nuclear and cytosolic fractions. (F) HIF transcriptional activity was measured in HeyA8 cells transiently co-transfected with the 5HRE reporter system in conjunction with either constitutively active HIF1α or HIF2α plasmids. Cells were then treated ± metformin (1mM, 24h) (n=3). Data represent mean value ± S.D. * p

    Journal: Molecular cancer research : MCR

    Article Title: SPHK1 is a novel target of metformin in ovarian cancer

    doi: 10.1158/1541-7786.MCR-18-0409

    Figure Lengend Snippet: Metformin inhibits hypoxia-induced expression of SPHK1 in OvCa. (A) SPHK1 protein expression was assessed by immunoblot in OVCAR5, Kuramochi and TYKnu cells overexpressing constitutively stable HIF1α with P402A/P564A (left) or HIF2α with P405A/P531A (right) mutations (n=3). (B) SPHK1 protein expression was analyzed by immunoblot after metformin treatment (1mM, 72h) in OVCAR5 and Kuramochi cells overexpressing constitutively stable HIF1α with activating substitution P402A/P564A (left) or HIF2α with P405A/P531A (right) (n=3). (C) HIF1α, HIF2α and SPHK1 protein expression were assessed by immunoblot in response to hypoxia (1% O 2 , 5% CO 2 ) with metformin concurrent treatment (1mM, 48h) (n=3). (D) HIF1α and HIF2α protein expression and localization in OVCAR5 cells was measured by immunoblot following cellular fractionation. HIF expression was stabilized using CoCl 2 (100μM) and concurrently treated with metformin (1mM, 24h). Histone H3 and βTubulin were used to normalize nuclear and cytosolic localization, respectively (n=3). (E) HIF1α and HIF2α protein expression and localization in OVCAR5 cells was measured by immunoblot following cellular fractionation following transfection with HIF1α or HIF2α constitutively stable mutants and subsequent treatment with metformin (1mM, 24h). Histone H3 and βTubulin were used to confirm nuclear and cytosolic fractions. (F) HIF transcriptional activity was measured in HeyA8 cells transiently co-transfected with the 5HRE reporter system in conjunction with either constitutively active HIF1α or HIF2α plasmids. Cells were then treated ± metformin (1mM, 24h) (n=3). Data represent mean value ± S.D. * p

    Article Snippet: Together, these data support the S1P pathway and SPHK1 in particular, as potential targets for OvCa treatment.

    Techniques: Expressing, Cell Fractionation, Transfection, Activity Assay

    SPHK1 expression is associated with metformin sensitivity in OvCa cells. (A) MTT viability assay of six OvCa cell lines analyzing sensitivity to metformin using a dose range up to 40mM in 5mM glucose DMEM (n=6). SPHK1 and SGPL1 mRNA (B) and protein (C) expression was assessed in multiple OvCa cell lines (n=3). (D) MTT viability assay IC 50 values for metformin (mM) in TYKnu, OVCAR5 and Kuramochi cells overexpressing SPHK1 or control vector (n=6). (E) Expression of cleaved caspase 3 was assessed through immunoblot in OVCAR5 and Kuramochi cells overexpressing SPHK1 or control vector with concurrent treatment with metformin (1mM, 48h) (n=3). (F) MTT viability assay IC 50 values for metformin (mM) in TYKnu cells with transient knockdown of SPHK1 using siRNA, in either normoxia or hypoxia (1% O 2 , 5% CO 2 ) (n=6). (G) Knockdown of SPHK1 mRNA expression was confirmed by qRT-PCR following knockdown using siRNA in HeyA8 cells (n=3). (H) Western blot of SPHK1 and cleaved caspase 3 in TYKnu cells following knockdown of SPHK1 using siRNA, with or without metformin treatment (1mM, 48h) (n=3). Data represent mean value ± S.D.

    Journal: Molecular cancer research : MCR

    Article Title: SPHK1 is a novel target of metformin in ovarian cancer

    doi: 10.1158/1541-7786.MCR-18-0409

    Figure Lengend Snippet: SPHK1 expression is associated with metformin sensitivity in OvCa cells. (A) MTT viability assay of six OvCa cell lines analyzing sensitivity to metformin using a dose range up to 40mM in 5mM glucose DMEM (n=6). SPHK1 and SGPL1 mRNA (B) and protein (C) expression was assessed in multiple OvCa cell lines (n=3). (D) MTT viability assay IC 50 values for metformin (mM) in TYKnu, OVCAR5 and Kuramochi cells overexpressing SPHK1 or control vector (n=6). (E) Expression of cleaved caspase 3 was assessed through immunoblot in OVCAR5 and Kuramochi cells overexpressing SPHK1 or control vector with concurrent treatment with metformin (1mM, 48h) (n=3). (F) MTT viability assay IC 50 values for metformin (mM) in TYKnu cells with transient knockdown of SPHK1 using siRNA, in either normoxia or hypoxia (1% O 2 , 5% CO 2 ) (n=6). (G) Knockdown of SPHK1 mRNA expression was confirmed by qRT-PCR following knockdown using siRNA in HeyA8 cells (n=3). (H) Western blot of SPHK1 and cleaved caspase 3 in TYKnu cells following knockdown of SPHK1 using siRNA, with or without metformin treatment (1mM, 48h) (n=3). Data represent mean value ± S.D.

    Article Snippet: Together, these data support the S1P pathway and SPHK1 in particular, as potential targets for OvCa treatment.

    Techniques: Expressing, MTT Assay, Viability Assay, Plasmid Preparation, Quantitative RT-PCR, Western Blot

    MiR-124 directly targets SphK1 and inhibits the migration and invasion of ovarian cancer cells. (A) Wound healing assay of SKOV3-ip and HO8910pm cells transfected with NC or miR-124 mimics. Representative pictures are shown at 0 and 24 h after the wound was made. (B) Transwell assay of SKOV3-ip and HO8910pm cells transfected with NC or miR-124 mimics. (C) Matrigel invasion assay of SKOV3-ip and HO8910pm cells transfected with miR-124 mimics or NC. (D) Expression of endogenous SphK1 in SKOV3-ip cells at 48 h and 72 h post transfection of miR-124 mimics or NC. β-actin was loaded as an internal control. (E) Diagram of the SphK1-3’-UTR with potential binding-sites for miR-124. (F) Relative luciferase activity of reporters including wild-type or mutant SphK1 3’-UTR co-transfected with NC or miR-124 mimics. ** and *** indicate significant difference at p

    Journal: Journal of Ovarian Research

    Article Title: MiR-124 inhibits the migration and invasion of ovarian cancer cells by targeting SphK1

    doi: 10.1186/1757-2215-6-84

    Figure Lengend Snippet: MiR-124 directly targets SphK1 and inhibits the migration and invasion of ovarian cancer cells. (A) Wound healing assay of SKOV3-ip and HO8910pm cells transfected with NC or miR-124 mimics. Representative pictures are shown at 0 and 24 h after the wound was made. (B) Transwell assay of SKOV3-ip and HO8910pm cells transfected with NC or miR-124 mimics. (C) Matrigel invasion assay of SKOV3-ip and HO8910pm cells transfected with miR-124 mimics or NC. (D) Expression of endogenous SphK1 in SKOV3-ip cells at 48 h and 72 h post transfection of miR-124 mimics or NC. β-actin was loaded as an internal control. (E) Diagram of the SphK1-3’-UTR with potential binding-sites for miR-124. (F) Relative luciferase activity of reporters including wild-type or mutant SphK1 3’-UTR co-transfected with NC or miR-124 mimics. ** and *** indicate significant difference at p

    Article Snippet: The results are consistent with previous research [ ], indicating that SphK1 is a predicted target of miR-124, and the inhibitory effect of miR-124 is due to direct interaction with the putative binding site of the 3’-UTR of SphK1.

    Techniques: Migration, Wound Healing Assay, Transfection, Transwell Assay, Invasion Assay, Expressing, Binding Assay, Luciferase, Activity Assay, Mutagenesis

    SphK1 knockdown results in inhibition of migration and invasion in ovarian cancer cells. (A) SphK1 detection in SKOV3-ip and HO8910pm cells transfected with NC or pooled si-SphK1. (B) Wound healing assay of SKOV3-ip and HO8910pm cells after transfected with NC or pooled si-SphK1. (C) Transwell assay of SKOV3-ip and HO8910pm cells subjected to NC or pooled si-SphK1. (D) Matrigel invasion assay of SKOV3-ip and HO8910pm cells after transfection with NC or pooled si-SphK1. Symbols ** and *** represent significance at p

    Journal: Journal of Ovarian Research

    Article Title: MiR-124 inhibits the migration and invasion of ovarian cancer cells by targeting SphK1

    doi: 10.1186/1757-2215-6-84

    Figure Lengend Snippet: SphK1 knockdown results in inhibition of migration and invasion in ovarian cancer cells. (A) SphK1 detection in SKOV3-ip and HO8910pm cells transfected with NC or pooled si-SphK1. (B) Wound healing assay of SKOV3-ip and HO8910pm cells after transfected with NC or pooled si-SphK1. (C) Transwell assay of SKOV3-ip and HO8910pm cells subjected to NC or pooled si-SphK1. (D) Matrigel invasion assay of SKOV3-ip and HO8910pm cells after transfection with NC or pooled si-SphK1. Symbols ** and *** represent significance at p

    Article Snippet: The results are consistent with previous research [ ], indicating that SphK1 is a predicted target of miR-124, and the inhibitory effect of miR-124 is due to direct interaction with the putative binding site of the 3’-UTR of SphK1.

    Techniques: Inhibition, Migration, Transfection, Wound Healing Assay, Transwell Assay, Invasion Assay

    Expression of SphK1 reversed the miR-124-induced inhibition of cellular migration and invasion. (A) The transfection of pcDNA3.1 (−)–SphK1 restored the expression of Sphk1 in SKOV3-ip cells even with miR-124 co-transfection. (B) Transwell assay and (C) Matrigel invasion assay of SKOV3-ip cells after co-transfection of NC or miR-124 mimics together with either pcDNA3.1 (−)-vector or pcDNA3.1 (−)-SphK1. ***denotes statistical significance at p

    Journal: Journal of Ovarian Research

    Article Title: MiR-124 inhibits the migration and invasion of ovarian cancer cells by targeting SphK1

    doi: 10.1186/1757-2215-6-84

    Figure Lengend Snippet: Expression of SphK1 reversed the miR-124-induced inhibition of cellular migration and invasion. (A) The transfection of pcDNA3.1 (−)–SphK1 restored the expression of Sphk1 in SKOV3-ip cells even with miR-124 co-transfection. (B) Transwell assay and (C) Matrigel invasion assay of SKOV3-ip cells after co-transfection of NC or miR-124 mimics together with either pcDNA3.1 (−)-vector or pcDNA3.1 (−)-SphK1. ***denotes statistical significance at p

    Article Snippet: The results are consistent with previous research [ ], indicating that SphK1 is a predicted target of miR-124, and the inhibitory effect of miR-124 is due to direct interaction with the putative binding site of the 3’-UTR of SphK1.

    Techniques: Expressing, Inhibition, Migration, Transfection, Cotransfection, Transwell Assay, Invasion Assay, Plasmid Preparation

    Impact of tamoxifen and LCL204 on AC and Sphk1 expression and Sphk1 activity in AML cell lines. (A) Tamoxifen dose-response in KG-1 cells. KG-1 cells were seeded in 6-well plates, 5 × 10 6 /well, in media containing 5% FBS. After 60 min, cells were

    Journal: Biochimica et biophysica acta

    Article Title: Modification of sphingolipid metabolism by tamoxifen and N-desmethyltamoxifen in acute myelogenous leukemia – Impact on enzyme activity and response to cytotoxics

    doi: 10.1016/j.bbalip.2015.03.001

    Figure Lengend Snippet: Impact of tamoxifen and LCL204 on AC and Sphk1 expression and Sphk1 activity in AML cell lines. (A) Tamoxifen dose-response in KG-1 cells. KG-1 cells were seeded in 6-well plates, 5 × 10 6 /well, in media containing 5% FBS. After 60 min, cells were

    Article Snippet: Sphk1 activity was quantified by using a commercial Sphingosine Kinase Activity Assay Kit (Echelon Biosciences, Salt Lake City, UT) as the manufacturer instructed.

    Techniques: Expressing, Activity Assay