angiogenic inducer 61 cyr61 Search Results


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    Cell Signaling Technology Inc angiogenic inducer 61 cyr61
    Angiogenic Inducer 61 Cyr61, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology angiogenic inducer 61 cyr61
    Angiogenic Inducer 61 Cyr61, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam angiogenic inducer 61 cyr61 antibodies
    Expression levels of fibulin-4, <t>CYR61</t> and ALP in human osteosarcoma tissues. (A) Fibulin-4 expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (B) ALP expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (C) CYR61 expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (D) According to the Pearson's product-moment correlation coefficient, the expression of fibulin-4 positively correlated with CYR61 and ALP, using photodensities detected by software Image-Pro Plus 6.0. (Magnification, ×200).
    Angiogenic Inducer 61 Cyr61 Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cusabio angiogenic inducer 61 cyr61 elisa kit
    RNA-seq on placentas from mice exposed to NaAsO 2 . (A–G) All pregnant mice except controls were exposed to NaAsO 2 ( 15 mg / L ) by drinking water throughout pregnancy. All pregnant mice were sacrificed on GD 18. (A) Volcano plot of differentially expressed genes in mouse placentas. A total of 743 genes (red triangle) were up-regulated, and 1,889 genes (green dot) were down-regulated by at least 1.5-fold with a significance level ( p < 0.05 ). (B) KEGG pathway analysis. (C) Top 100 genes were screened based on the p -value ranking of all differential genes and then we screened 40 candidate genes among them, with the average FPKM > 10 of all samples as the filter condition. RNA-seq of mouse placentae from three dams were analyzed per group. (D) Validation of trophoblasts development-related differentially expressed genes by real-time RT-PCR ( n = 6 mice/group). (E) Fold differences in differentially expressed genes between RNA-seq and real-time RT-PCR. (F) Placental <t>CYR61</t> was detected by immunoblotting ( n = 3 mice/group, repeated three times). (G) Representative pictures of CYR61-positive cells in labyrinth zone and junctional zone. Scale bars, 20 μ m ( n = 6 mice/group). (H–J) HTR-8/SVneo cells were exposed to NaAsO 2 ( 2 μ M ). (H) Cyr61 mRNA was measured by real-time RT-PCR ( n = 6 samples/group). (I) <t>CYR61</t> <t>protein</t> was detected by immunoblotting ( n = 3 samples/group, repeated three times). (J) CYR61-positive cells were stained with by immunofluorescence ( n = 3 samples/group). All data are expressed as mean ± SEM . The numeric data are shown in Table S8. RNA-seq data are reported in an Excel sheet and uploaded to the National Center for Biotechnology Information (GSE222092). Unpaired two-tailed Student’s t -test was used for D–F, H, I. * p < 0.05 , ** p < 0.01 . Note: As, arsenic; As-H, 15 mg / L NaAsO 2 ; Ceacam 3/13, carcinoembryonic antigen-related cell adhesion molecule 3/13; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; FPKM, fragments per kilobase per million; Not DEG, not differential gene expression; RNA-seq, RNA sequencing; RT-PCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean; Sparcl1, SPARC-like protein 1; TGF- β 2 , transforming growth factor β 2 ; μ M , μ mol / L .
    Angiogenic Inducer 61 Cyr61 Elisa Kit, supplied by Cusabio, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam cysteine rich angiogenic inducer 61 cyr61
    Primer Sequences for RT-qPCR
    Cysteine Rich Angiogenic Inducer 61 Cyr61, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Expression levels of fibulin-4, CYR61 and ALP in human osteosarcoma tissues. (A) Fibulin-4 expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (B) ALP expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (C) CYR61 expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (D) According to the Pearson's product-moment correlation coefficient, the expression of fibulin-4 positively correlated with CYR61 and ALP, using photodensities detected by software Image-Pro Plus 6.0. (Magnification, ×200).

    Journal: International Journal of Oncology

    Article Title: Fibulin-4 promotes osteosarcoma invasion and metastasis by inducing epithelial to mesenchymal transition via the PI3K/Akt/mTOR pathway

    doi: 10.3892/ijo.2017.3921

    Figure Lengend Snippet: Expression levels of fibulin-4, CYR61 and ALP in human osteosarcoma tissues. (A) Fibulin-4 expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (B) ALP expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (C) CYR61 expression levels in osteosarcoma tissue, fibrous dysplasia tissue and normal tissue were measured by IHC. (D) According to the Pearson's product-moment correlation coefficient, the expression of fibulin-4 positively correlated with CYR61 and ALP, using photodensities detected by software Image-Pro Plus 6.0. (Magnification, ×200).

    Article Snippet: All sections and coverslips were treated with 3% hydrogen peroxide (H 2 O 2 ) and goat serum for 30 min sequentially, to block endogenous peroxidase and the non-specific binding sites, and then incubated with rabbit anti-human fibulin-4, alkaline phosphatase (ALP), and cysteine-rich angiogenic inducer 61 (CYR61) antibodies (ab125073, ab108337, ab24448; Abcam) at working dilutions of 1:200 overnight at 4°C.

    Techniques: Expressing, Software

    Effects of fibulin-4 knockdown and overexpression on EMT genes, CYR61 and ALP, which were correlated to tumor progression. After lentivirus transfections, EMT markers, including E-cadherin, N-cadherin, vimentin, Snail, Slug and Twist were measured by (A) western blotting and (B) real-time q-RT-PCR in the lentivirus transfection systems of the differently invasive osteosarcoma cell subclones. (C) EMT markers (E-cadherin, N-cadherin, vimentin, Snail, Slug and Twist), CYR61 and ALP were detected by western blotting in fibulin-4 shRNA-infected HOS cells and pLVX-fibulin-4-infected U-2OS cells. Fibulin-4 knockdown significantly increased the expression of E-cadherin, and decreased the expression of N-cadherin, vimentin, Snail, Slug and Twist; whereas fibulin-4 upregulation decreased the expression of E-cadherin, and increased the expression of N-cadherin, vimentin, Snail, Slug and Twist. CYR61 and ALP were downregulated following fibulin-4 shRNA infection, in association with the knockdown of fibulin-4 expression, moreover, with fibulin-4 upregulation, CYR61 and ALP were increased following pLVX-fibulin-4 infection. * P<0.05.

    Journal: International Journal of Oncology

    Article Title: Fibulin-4 promotes osteosarcoma invasion and metastasis by inducing epithelial to mesenchymal transition via the PI3K/Akt/mTOR pathway

    doi: 10.3892/ijo.2017.3921

    Figure Lengend Snippet: Effects of fibulin-4 knockdown and overexpression on EMT genes, CYR61 and ALP, which were correlated to tumor progression. After lentivirus transfections, EMT markers, including E-cadherin, N-cadherin, vimentin, Snail, Slug and Twist were measured by (A) western blotting and (B) real-time q-RT-PCR in the lentivirus transfection systems of the differently invasive osteosarcoma cell subclones. (C) EMT markers (E-cadherin, N-cadherin, vimentin, Snail, Slug and Twist), CYR61 and ALP were detected by western blotting in fibulin-4 shRNA-infected HOS cells and pLVX-fibulin-4-infected U-2OS cells. Fibulin-4 knockdown significantly increased the expression of E-cadherin, and decreased the expression of N-cadherin, vimentin, Snail, Slug and Twist; whereas fibulin-4 upregulation decreased the expression of E-cadherin, and increased the expression of N-cadherin, vimentin, Snail, Slug and Twist. CYR61 and ALP were downregulated following fibulin-4 shRNA infection, in association with the knockdown of fibulin-4 expression, moreover, with fibulin-4 upregulation, CYR61 and ALP were increased following pLVX-fibulin-4 infection. * P<0.05.

    Article Snippet: All sections and coverslips were treated with 3% hydrogen peroxide (H 2 O 2 ) and goat serum for 30 min sequentially, to block endogenous peroxidase and the non-specific binding sites, and then incubated with rabbit anti-human fibulin-4, alkaline phosphatase (ALP), and cysteine-rich angiogenic inducer 61 (CYR61) antibodies (ab125073, ab108337, ab24448; Abcam) at working dilutions of 1:200 overnight at 4°C.

    Techniques: Over Expression, Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction, shRNA, Infection, Expressing

    RNA-seq on placentas from mice exposed to NaAsO 2 . (A–G) All pregnant mice except controls were exposed to NaAsO 2 ( 15 mg / L ) by drinking water throughout pregnancy. All pregnant mice were sacrificed on GD 18. (A) Volcano plot of differentially expressed genes in mouse placentas. A total of 743 genes (red triangle) were up-regulated, and 1,889 genes (green dot) were down-regulated by at least 1.5-fold with a significance level ( p < 0.05 ). (B) KEGG pathway analysis. (C) Top 100 genes were screened based on the p -value ranking of all differential genes and then we screened 40 candidate genes among them, with the average FPKM > 10 of all samples as the filter condition. RNA-seq of mouse placentae from three dams were analyzed per group. (D) Validation of trophoblasts development-related differentially expressed genes by real-time RT-PCR ( n = 6 mice/group). (E) Fold differences in differentially expressed genes between RNA-seq and real-time RT-PCR. (F) Placental CYR61 was detected by immunoblotting ( n = 3 mice/group, repeated three times). (G) Representative pictures of CYR61-positive cells in labyrinth zone and junctional zone. Scale bars, 20 μ m ( n = 6 mice/group). (H–J) HTR-8/SVneo cells were exposed to NaAsO 2 ( 2 μ M ). (H) Cyr61 mRNA was measured by real-time RT-PCR ( n = 6 samples/group). (I) CYR61 protein was detected by immunoblotting ( n = 3 samples/group, repeated three times). (J) CYR61-positive cells were stained with by immunofluorescence ( n = 3 samples/group). All data are expressed as mean ± SEM . The numeric data are shown in Table S8. RNA-seq data are reported in an Excel sheet and uploaded to the National Center for Biotechnology Information (GSE222092). Unpaired two-tailed Student’s t -test was used for D–F, H, I. * p < 0.05 , ** p < 0.01 . Note: As, arsenic; As-H, 15 mg / L NaAsO 2 ; Ceacam 3/13, carcinoembryonic antigen-related cell adhesion molecule 3/13; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; FPKM, fragments per kilobase per million; Not DEG, not differential gene expression; RNA-seq, RNA sequencing; RT-PCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean; Sparcl1, SPARC-like protein 1; TGF- β 2 , transforming growth factor β 2 ; μ M , μ mol / L .

    Journal: Environmental Health Perspectives

    Article Title: Effects of Gestational Arsenic Exposures on Placental and Fetal Development in Mice: The Role of Cyr61 m 6 A

    doi: 10.1289/EHP12207

    Figure Lengend Snippet: RNA-seq on placentas from mice exposed to NaAsO 2 . (A–G) All pregnant mice except controls were exposed to NaAsO 2 ( 15 mg / L ) by drinking water throughout pregnancy. All pregnant mice were sacrificed on GD 18. (A) Volcano plot of differentially expressed genes in mouse placentas. A total of 743 genes (red triangle) were up-regulated, and 1,889 genes (green dot) were down-regulated by at least 1.5-fold with a significance level ( p < 0.05 ). (B) KEGG pathway analysis. (C) Top 100 genes were screened based on the p -value ranking of all differential genes and then we screened 40 candidate genes among them, with the average FPKM > 10 of all samples as the filter condition. RNA-seq of mouse placentae from three dams were analyzed per group. (D) Validation of trophoblasts development-related differentially expressed genes by real-time RT-PCR ( n = 6 mice/group). (E) Fold differences in differentially expressed genes between RNA-seq and real-time RT-PCR. (F) Placental CYR61 was detected by immunoblotting ( n = 3 mice/group, repeated three times). (G) Representative pictures of CYR61-positive cells in labyrinth zone and junctional zone. Scale bars, 20 μ m ( n = 6 mice/group). (H–J) HTR-8/SVneo cells were exposed to NaAsO 2 ( 2 μ M ). (H) Cyr61 mRNA was measured by real-time RT-PCR ( n = 6 samples/group). (I) CYR61 protein was detected by immunoblotting ( n = 3 samples/group, repeated three times). (J) CYR61-positive cells were stained with by immunofluorescence ( n = 3 samples/group). All data are expressed as mean ± SEM . The numeric data are shown in Table S8. RNA-seq data are reported in an Excel sheet and uploaded to the National Center for Biotechnology Information (GSE222092). Unpaired two-tailed Student’s t -test was used for D–F, H, I. * p < 0.05 , ** p < 0.01 . Note: As, arsenic; As-H, 15 mg / L NaAsO 2 ; Ceacam 3/13, carcinoembryonic antigen-related cell adhesion molecule 3/13; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; FPKM, fragments per kilobase per million; Not DEG, not differential gene expression; RNA-seq, RNA sequencing; RT-PCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean; Sparcl1, SPARC-like protein 1; TGF- β 2 , transforming growth factor β 2 ; μ M , μ mol / L .

    Article Snippet: The cysteine-rich angiogenic inducer 61 (CYR61) ELISA kit (#CSB-E13884h) was purchased from Cusabio.

    Techniques: RNA Sequencing Assay, Quantitative RT-PCR, Western Blot, Staining, Immunofluorescence, Two Tailed Test, Expressing, Reverse Transcription Polymerase Chain Reaction

    The role of CYR61 in induced inhibition of migration and invasion in As-exposed human placental trophoblasts. HTR-8/SVneo cells were exposed to NaAsO 2 ( 2 μ M ). (A–D) HTR-8/SVneo cells were transfected with CYR61 overexpression plasmid and then exposed to NaAsO 2 ( 2 μ M ). (A) The efficiency of CYR61 overexpression was determined by real-time RT-PCR ( n = 6 samples/group). (B) Vimentin, N-cadherin, and MMP2 proteins were measured by immunoblotting ( n = 2 samples/group, repeated three times), and β -actin was used as a loading control. (C) Cell migration was determined by transwell migration assay ( n = 3 samples/group, repeated two times). (D) Cell invasion was measured by transwell invasion assay ( n = 3 samples/group, repeated 2 times). (E–H) HTR-8/SVneo cells were transfected with CYR61 knockdown plasmid and then exposed to NaAsO 2 ( 2 μ M ). (E) The efficiency of CYR61 knockdown was determined by real-time RT-PCR ( n = 6 samples/group). (F) Vimentin, N-cadherin, and MMP2 proteins were measured by immunoblotting ( n = 2 samples/group, repeated three times); β -actin was used as a loading control. (G) Cell migration was determined by transwell migration assay ( n = 3 samples/group, repeated two times). (H) Cell invasion was measured by transwell invasion assay ( n = 3 samples/group, repeated 2 times). Scale bars, 200 μ m . All data were expressed as mean ± SEM . The numeric data are shown in Table S8. Unpaired two-tailed Student’s t -test was used for A and E. When ANOVA showed p < 0.05 , the Student-Neuman-Keuls (SNK) multiple comparisons tests were used for B–D and F–H. * p < 0.05 , ** p < 0.01 . Note: ANOVA, analysis of variance; As, arsenic; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; CYR61 OE, CYR61 overexpression; shCYR61, CYR61 shRNA; RT-PCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean; μ M , μ mol / L .

    Journal: Environmental Health Perspectives

    Article Title: Effects of Gestational Arsenic Exposures on Placental and Fetal Development in Mice: The Role of Cyr61 m 6 A

    doi: 10.1289/EHP12207

    Figure Lengend Snippet: The role of CYR61 in induced inhibition of migration and invasion in As-exposed human placental trophoblasts. HTR-8/SVneo cells were exposed to NaAsO 2 ( 2 μ M ). (A–D) HTR-8/SVneo cells were transfected with CYR61 overexpression plasmid and then exposed to NaAsO 2 ( 2 μ M ). (A) The efficiency of CYR61 overexpression was determined by real-time RT-PCR ( n = 6 samples/group). (B) Vimentin, N-cadherin, and MMP2 proteins were measured by immunoblotting ( n = 2 samples/group, repeated three times), and β -actin was used as a loading control. (C) Cell migration was determined by transwell migration assay ( n = 3 samples/group, repeated two times). (D) Cell invasion was measured by transwell invasion assay ( n = 3 samples/group, repeated 2 times). (E–H) HTR-8/SVneo cells were transfected with CYR61 knockdown plasmid and then exposed to NaAsO 2 ( 2 μ M ). (E) The efficiency of CYR61 knockdown was determined by real-time RT-PCR ( n = 6 samples/group). (F) Vimentin, N-cadherin, and MMP2 proteins were measured by immunoblotting ( n = 2 samples/group, repeated three times); β -actin was used as a loading control. (G) Cell migration was determined by transwell migration assay ( n = 3 samples/group, repeated two times). (H) Cell invasion was measured by transwell invasion assay ( n = 3 samples/group, repeated 2 times). Scale bars, 200 μ m . All data were expressed as mean ± SEM . The numeric data are shown in Table S8. Unpaired two-tailed Student’s t -test was used for A and E. When ANOVA showed p < 0.05 , the Student-Neuman-Keuls (SNK) multiple comparisons tests were used for B–D and F–H. * p < 0.05 , ** p < 0.01 . Note: ANOVA, analysis of variance; As, arsenic; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; CYR61 OE, CYR61 overexpression; shCYR61, CYR61 shRNA; RT-PCR, reverse transcription polymerase chain reaction; SEM, standard error of the mean; μ M , μ mol / L .

    Article Snippet: The cysteine-rich angiogenic inducer 61 (CYR61) ELISA kit (#CSB-E13884h) was purchased from Cusabio.

    Techniques: Inhibition, Migration, Transfection, Over Expression, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Transwell Migration Assay, Transwell Invasion Assay, Two Tailed Test, shRNA, Reverse Transcription Polymerase Chain Reaction

    Effect of m 6 A modification on Cyr61 mRNA stability and CYR61 protein in As-exposed human placental trophoblasts. HTR-8/SVneo cells were exposed to NaAsO 2 ( 2 μ M ). (A) HTR8/SVneo cells were treated with actinomycin D ( 5 μ g / mL ), followed by detection of Cyr61 mRNA at indicated times ( n = 3 samples/group, repeated two times). (B) The m 6 A content was measured by EpiQuik m 6 A methylation quantification kit ( n = 5 samples/group, repeated two times). (C) Schematic representation of the position of m 6 A motifs with CYR61 transcript. (D) The m 6 A -modified Cyr61 levels were measured by MeRIP-qPCR ( n = 3 samples/group). (E) Schematic representation of the interaction between Cyr61 mRNA and IGF2BP1/2/3 proteins. (F) The interaction between Cyr61 mRNA and IGF2BP2 proteins was detected by RIP-qPCR ( n = 3 samples/group). (G) The expression of m 6 A methyltransferases and demethylases was measured by real-time RT-PCR ( n = 6 samples/group). (H) The m 6 A methylase activity was measured by Epigenase m 6 A methylase activity/inhibition assay kit ( n = 6 samples/group). (I) A schematic model of the reversible m 6 A methylation. SAM is not only a methyl donor for m 6 A methylation but also a cofactor for catalytic domain of Mettl3. (J) Intracellular SAM content was measured by LC-MS/MS ( n = 6 samples/group). All data were expressed as mean ± SEM . The numeric data are shown in Table S8. Unpaired two-tailed Student’s t -test was used for A, B, D, F, G, H, and J. * p < 0.05 , ** p < 0.01 . Note: ActD, actinomycin D; Alkbh5, a-ketoglutarate-dependent dioxygenase alkB homolog 5; ANOVA, analysis of variance; As, arsenic; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; FTO, fat-mass obesity-associated protein; IGF2BP1/2/3, insulin-like growth factor 2 mRNA-binding protein one-half/3; LC-MS/MS, liquid chromatography–tandem mass spectrometry; m 6 A , N 6 -methyladenosine ; MeRIP-qPCR, methylated RNA immunoprecipitation–qPCR; Mettl3/14, methyltransferase-like 3/14; Rbm15/15b, RNA-binding motif protein 15/15b; RIP-qPCR, RNA-binding protein immunoprecipitation–qPCR; RT-PCR, reverse transcription polymerase chain reaction; SAH, s-adenosyl-l-homocysteine; SAM, s-adenosylmethionine; SEM, standard error of the mean; μ M , μ mol / L ; WTAP, wilms tumor 1-associating protein.

    Journal: Environmental Health Perspectives

    Article Title: Effects of Gestational Arsenic Exposures on Placental and Fetal Development in Mice: The Role of Cyr61 m 6 A

    doi: 10.1289/EHP12207

    Figure Lengend Snippet: Effect of m 6 A modification on Cyr61 mRNA stability and CYR61 protein in As-exposed human placental trophoblasts. HTR-8/SVneo cells were exposed to NaAsO 2 ( 2 μ M ). (A) HTR8/SVneo cells were treated with actinomycin D ( 5 μ g / mL ), followed by detection of Cyr61 mRNA at indicated times ( n = 3 samples/group, repeated two times). (B) The m 6 A content was measured by EpiQuik m 6 A methylation quantification kit ( n = 5 samples/group, repeated two times). (C) Schematic representation of the position of m 6 A motifs with CYR61 transcript. (D) The m 6 A -modified Cyr61 levels were measured by MeRIP-qPCR ( n = 3 samples/group). (E) Schematic representation of the interaction between Cyr61 mRNA and IGF2BP1/2/3 proteins. (F) The interaction between Cyr61 mRNA and IGF2BP2 proteins was detected by RIP-qPCR ( n = 3 samples/group). (G) The expression of m 6 A methyltransferases and demethylases was measured by real-time RT-PCR ( n = 6 samples/group). (H) The m 6 A methylase activity was measured by Epigenase m 6 A methylase activity/inhibition assay kit ( n = 6 samples/group). (I) A schematic model of the reversible m 6 A methylation. SAM is not only a methyl donor for m 6 A methylation but also a cofactor for catalytic domain of Mettl3. (J) Intracellular SAM content was measured by LC-MS/MS ( n = 6 samples/group). All data were expressed as mean ± SEM . The numeric data are shown in Table S8. Unpaired two-tailed Student’s t -test was used for A, B, D, F, G, H, and J. * p < 0.05 , ** p < 0.01 . Note: ActD, actinomycin D; Alkbh5, a-ketoglutarate-dependent dioxygenase alkB homolog 5; ANOVA, analysis of variance; As, arsenic; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; FTO, fat-mass obesity-associated protein; IGF2BP1/2/3, insulin-like growth factor 2 mRNA-binding protein one-half/3; LC-MS/MS, liquid chromatography–tandem mass spectrometry; m 6 A , N 6 -methyladenosine ; MeRIP-qPCR, methylated RNA immunoprecipitation–qPCR; Mettl3/14, methyltransferase-like 3/14; Rbm15/15b, RNA-binding motif protein 15/15b; RIP-qPCR, RNA-binding protein immunoprecipitation–qPCR; RT-PCR, reverse transcription polymerase chain reaction; SAH, s-adenosyl-l-homocysteine; SAM, s-adenosylmethionine; SEM, standard error of the mean; μ M , μ mol / L ; WTAP, wilms tumor 1-associating protein.

    Article Snippet: The cysteine-rich angiogenic inducer 61 (CYR61) ELISA kit (#CSB-E13884h) was purchased from Cusabio.

    Techniques: Modification, Methylation, Expressing, Quantitative RT-PCR, Activity Assay, Inhibition, Liquid Chromatography with Mass Spectroscopy, Two Tailed Test, Binding Assay, Liquid Chromatography, Mass Spectrometry, Immunoprecipitation, RNA Binding Assay, Reverse Transcription Polymerase Chain Reaction, Wilms Tumor Assay

    Effect of As3MT knockdown on Cyr61 m 6 A modification in As-exposed human placental trophoblasts. HTR8/SVneo cells were exposed to 2 μ M NaAsO 2 in presence or absence of As3MT-shRNA (A) The efficiency of As3MT knockdown was determined by real-time RT-PCR ( n = 6 samples/group). (B) Intracellular SAM content was measured by LC-MS/MS ( n = 6 samples/group). (C) The m 6 A methylase activity was measured by Epigenase m 6 A methylase activity/inhibition assay kit ( n = 6 samples/group). (D and E) HTR8/SVneo cells were treated with actinomycin D ( 5 μ g / mL ), followed by detection of Cyr61 mRNA at indicated times ( n = 3 samples/group, repeated two times). (F) CYR61, Vimentin, N-cadherin, and MMP2 proteins were measured by immunoblotting ( n = 3 samples/group). (G) Cell migration was determined by transwell migration assay ( n = 3 samples/group, repeated two times). (H) Cell invasion was measured by transwell invasion assay ( n = 3 samples/group, repeated two times). Scale bars, 200 μ m . All data were expressed as mean ± SEM . The numeric data are shown in Table S8. When ANOVA showed p < 0.05 , the Student-Neuman-Keuls (SNK) multiple comparisons tests were used for A–H. Asterisks indicate comparison with the control group. Hashtags indicate comparison with the As group. * p and # p < 0.05 , ** p and ## p < 0.01 . Note: ActD, actinomycin D; ANOVA, analysis of variance; As, arsenic; As3MT, arsenite methyltransferase; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; LC-MS/MS, liquid chromatography–tandem mass spectrometry; MMP2, matrix metalloproteinases 2; RT-PCR, reverse transcription polymerase chain reaction; SAM, s-adenosylmethionine; shAs3MT 1#, As3MT shRNA 1#; shAs3MT 2#, As3MT shRNA 2#; SEM, standard error of the mean; μ M , μ mol / L .

    Journal: Environmental Health Perspectives

    Article Title: Effects of Gestational Arsenic Exposures on Placental and Fetal Development in Mice: The Role of Cyr61 m 6 A

    doi: 10.1289/EHP12207

    Figure Lengend Snippet: Effect of As3MT knockdown on Cyr61 m 6 A modification in As-exposed human placental trophoblasts. HTR8/SVneo cells were exposed to 2 μ M NaAsO 2 in presence or absence of As3MT-shRNA (A) The efficiency of As3MT knockdown was determined by real-time RT-PCR ( n = 6 samples/group). (B) Intracellular SAM content was measured by LC-MS/MS ( n = 6 samples/group). (C) The m 6 A methylase activity was measured by Epigenase m 6 A methylase activity/inhibition assay kit ( n = 6 samples/group). (D and E) HTR8/SVneo cells were treated with actinomycin D ( 5 μ g / mL ), followed by detection of Cyr61 mRNA at indicated times ( n = 3 samples/group, repeated two times). (F) CYR61, Vimentin, N-cadherin, and MMP2 proteins were measured by immunoblotting ( n = 3 samples/group). (G) Cell migration was determined by transwell migration assay ( n = 3 samples/group, repeated two times). (H) Cell invasion was measured by transwell invasion assay ( n = 3 samples/group, repeated two times). Scale bars, 200 μ m . All data were expressed as mean ± SEM . The numeric data are shown in Table S8. When ANOVA showed p < 0.05 , the Student-Neuman-Keuls (SNK) multiple comparisons tests were used for A–H. Asterisks indicate comparison with the control group. Hashtags indicate comparison with the As group. * p and # p < 0.05 , ** p and ## p < 0.01 . Note: ActD, actinomycin D; ANOVA, analysis of variance; As, arsenic; As3MT, arsenite methyltransferase; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; LC-MS/MS, liquid chromatography–tandem mass spectrometry; MMP2, matrix metalloproteinases 2; RT-PCR, reverse transcription polymerase chain reaction; SAM, s-adenosylmethionine; shAs3MT 1#, As3MT shRNA 1#; shAs3MT 2#, As3MT shRNA 2#; SEM, standard error of the mean; μ M , μ mol / L .

    Article Snippet: The cysteine-rich angiogenic inducer 61 (CYR61) ELISA kit (#CSB-E13884h) was purchased from Cusabio.

    Techniques: Modification, shRNA, Quantitative RT-PCR, Liquid Chromatography with Mass Spectroscopy, Activity Assay, Inhibition, Western Blot, Migration, Transwell Migration Assay, Transwell Invasion Assay, Comparison, Liquid Chromatography, Mass Spectrometry, Reverse Transcription Polymerase Chain Reaction

    Effect of SAM supplementation on induced inhibition of Cyr61 m 6 A modification in As-exposed human placental trophoblasts. HTR8/SVneo cells were incubated with NaAsO 2 ( 2 μ M ) in presence or absence of SAM ( 10 μ M ). (A) Intracellular SAM content was measured by LC-MS/MS ( n = 6 samples/group). (B) The m 6 A methylase activity was measured by Epigenase m 6 A methylase activity/inhibition assay kit ( n = 6 samples/group). (C) HTR8/SVneo cells were treated with actinomycin D ( 5 μ g / mL ), followed by detection of Cyr61 mRNA at indicated times ( n = 3 samples/group, repeated two times). (D and E) CYR61, Vimentin, N-cadherin, and MMP2 proteins were determined by immunoblotting ( n = 3 samples/group, repeated three times). (F) Cell migration was determined by transwell migration assay ( n = 3 samples/group, repeated two times). (G) Cell invasion was measured by transwell invasion assay ( n = 6 samples/group, repeated two times). Scale bars, 200 μ m . All data were expressed as mean ± SEM . The numeric data are shown in Table S8. When ANOVA showed p < 0.05 , the Student-Neuman-Keuls (SNK) multiple comparisons tests were used for A–G. Asterisks indicate comparison with the control group. Hashtags indicate comparison with the As group. * p and # p < 0.05 , ** p and ## p < 0.01 . Note: ANOVA, analysis of variance; As, arsenic; ActD, actinomycin D; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; MMP2, matrix metalloproteinases 2; RT-PCR, reverse transcription polymerase chain reaction; SAM, s-adenosylmethionine; SEM, standard error of the mean; μ M , μ mol / L .

    Journal: Environmental Health Perspectives

    Article Title: Effects of Gestational Arsenic Exposures on Placental and Fetal Development in Mice: The Role of Cyr61 m 6 A

    doi: 10.1289/EHP12207

    Figure Lengend Snippet: Effect of SAM supplementation on induced inhibition of Cyr61 m 6 A modification in As-exposed human placental trophoblasts. HTR8/SVneo cells were incubated with NaAsO 2 ( 2 μ M ) in presence or absence of SAM ( 10 μ M ). (A) Intracellular SAM content was measured by LC-MS/MS ( n = 6 samples/group). (B) The m 6 A methylase activity was measured by Epigenase m 6 A methylase activity/inhibition assay kit ( n = 6 samples/group). (C) HTR8/SVneo cells were treated with actinomycin D ( 5 μ g / mL ), followed by detection of Cyr61 mRNA at indicated times ( n = 3 samples/group, repeated two times). (D and E) CYR61, Vimentin, N-cadherin, and MMP2 proteins were determined by immunoblotting ( n = 3 samples/group, repeated three times). (F) Cell migration was determined by transwell migration assay ( n = 3 samples/group, repeated two times). (G) Cell invasion was measured by transwell invasion assay ( n = 6 samples/group, repeated two times). Scale bars, 200 μ m . All data were expressed as mean ± SEM . The numeric data are shown in Table S8. When ANOVA showed p < 0.05 , the Student-Neuman-Keuls (SNK) multiple comparisons tests were used for A–G. Asterisks indicate comparison with the control group. Hashtags indicate comparison with the As group. * p and # p < 0.05 , ** p and ## p < 0.01 . Note: ANOVA, analysis of variance; As, arsenic; ActD, actinomycin D; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; MMP2, matrix metalloproteinases 2; RT-PCR, reverse transcription polymerase chain reaction; SAM, s-adenosylmethionine; SEM, standard error of the mean; μ M , μ mol / L .

    Article Snippet: The cysteine-rich angiogenic inducer 61 (CYR61) ELISA kit (#CSB-E13884h) was purchased from Cusabio.

    Techniques: Inhibition, Modification, Incubation, Liquid Chromatography with Mass Spectroscopy, Activity Assay, Western Blot, Migration, Transwell Migration Assay, Transwell Invasion Assay, Comparison, Reverse Transcription Polymerase Chain Reaction

    Effect of FA supplementation on placental development and fetal growth restriction in As-exposed mice. Pregnant mice were divided into four groups: CTRL, FA, As, and As + FA groups. In the As and As + FA groups, pregnant mice drank ultrapure water containing NaAsO 2 ( 15 mg / L ) throughout pregnancy. In FA and As + FA groups, pregnant mice were administered FA ( 150 μ g / kg ) by gavage daily throughout pregnancy. All pregnant mice were sacrificed on GD18. (A) Fetal weight ( n = 8 dams/group, 12–15 fetuses per dam). (B) Crown–rump length ( n = 8 dams/group, 12–15 fetuses per dam). (C) Placental weight ( n = 8 dams/group, 12–15 fetuses per dam). (D) Placental diameter ( n = 8 dams/group, 12–15 fetuses per dam). (E) Placental cross-section was stained with H&E. Scale bars, 500 μ m (left); 200 μ m (right) ( n = 6 mice/group). (F) The percentage of labyrinth zone area in the entire placenta area ( n = 6 mice/group). (G) The ratio of cross-sectional thickness of labyrinth zone to junctional zone ( n = 6 mice/group). (H) Placental SAM content was determined by LC-MS/MS ( n = 6 mice/group). (I) The m 6 A methylase activity was measured by Epigenase m 6 A methylase activity/inhibition assay kit ( n = 6 mice/group). (J) Placental CYR61 and MMP2 were measured by immunoblotting ( n = 3 mice/group, repeated two times). All data were expressed as mean ± SEM . The numeric data are shown in Table S8. When ANOVA showed p < 0.05 , the Student-Neuman-Keuls (SNK) multiple comparisons tests were used for A–D and F–J. * p < 0.05 , ** p < 0.01 . Note: ANOVA, analysis of variance; As, arsenic; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; FA, folic acid; H&E, hematoxylin and eosin; JZ, junctional zone; LC-MS/MS, liquid chromatography–tandem mass spectrometry; LZ, labyrinth zone; MMP2, matrix metalloproteinases 2; SAM, s-adenosylmethionine; SEM, standard error of the mean.

    Journal: Environmental Health Perspectives

    Article Title: Effects of Gestational Arsenic Exposures on Placental and Fetal Development in Mice: The Role of Cyr61 m 6 A

    doi: 10.1289/EHP12207

    Figure Lengend Snippet: Effect of FA supplementation on placental development and fetal growth restriction in As-exposed mice. Pregnant mice were divided into four groups: CTRL, FA, As, and As + FA groups. In the As and As + FA groups, pregnant mice drank ultrapure water containing NaAsO 2 ( 15 mg / L ) throughout pregnancy. In FA and As + FA groups, pregnant mice were administered FA ( 150 μ g / kg ) by gavage daily throughout pregnancy. All pregnant mice were sacrificed on GD18. (A) Fetal weight ( n = 8 dams/group, 12–15 fetuses per dam). (B) Crown–rump length ( n = 8 dams/group, 12–15 fetuses per dam). (C) Placental weight ( n = 8 dams/group, 12–15 fetuses per dam). (D) Placental diameter ( n = 8 dams/group, 12–15 fetuses per dam). (E) Placental cross-section was stained with H&E. Scale bars, 500 μ m (left); 200 μ m (right) ( n = 6 mice/group). (F) The percentage of labyrinth zone area in the entire placenta area ( n = 6 mice/group). (G) The ratio of cross-sectional thickness of labyrinth zone to junctional zone ( n = 6 mice/group). (H) Placental SAM content was determined by LC-MS/MS ( n = 6 mice/group). (I) The m 6 A methylase activity was measured by Epigenase m 6 A methylase activity/inhibition assay kit ( n = 6 mice/group). (J) Placental CYR61 and MMP2 were measured by immunoblotting ( n = 3 mice/group, repeated two times). All data were expressed as mean ± SEM . The numeric data are shown in Table S8. When ANOVA showed p < 0.05 , the Student-Neuman-Keuls (SNK) multiple comparisons tests were used for A–D and F–J. * p < 0.05 , ** p < 0.01 . Note: ANOVA, analysis of variance; As, arsenic; CTRL, control; CYR61, cysteine-rich angiogenic inducer 61; FA, folic acid; H&E, hematoxylin and eosin; JZ, junctional zone; LC-MS/MS, liquid chromatography–tandem mass spectrometry; LZ, labyrinth zone; MMP2, matrix metalloproteinases 2; SAM, s-adenosylmethionine; SEM, standard error of the mean.

    Article Snippet: The cysteine-rich angiogenic inducer 61 (CYR61) ELISA kit (#CSB-E13884h) was purchased from Cusabio.

    Techniques: Staining, Liquid Chromatography with Mass Spectroscopy, Activity Assay, Inhibition, Western Blot, Liquid Chromatography, Mass Spectrometry

    Based on a case–control study: association among maternal urinary As concentration, plasma CYR61 content, and fetal growth restriction. (A) Maternal urinary As concentration in AGA and SGA infants was determined by hydride generation-atomic fluorescence spectrometry ( n = 45 human sample/group). (B) Maternal plasma CYR61 content in AGA and SGA infants was measured by ELISA ( n = 45 human sample/group). (C) The correlation between maternal urinary As concentration and plasma CYR61 content in AGA infants ( n = 45 human sample). (D) The correlation between maternal urinary As concentration and plasma CYR61 content in SGA infants ( n = 45 human sample). (E) Placental CYR61-positive cells in SGA infants and AGA infants were stained by immunohistochemistry ( n = 18 human sample/group). Representative histology images were showed. Blue arrows indicate CYR61-positive cells. Scale bars, 20 μ m . (F) Placental vimentin, MMP2, and MMP9 were detected by immunoblotting ( n = 18 human sample/group). The data in A and B are expressed as mean ± SD . The data in A–D are represented by dots indicating individual values. The data in E and F are expressed as mean ± SEM . The numeric data are shown in Table S8. Two-tailed Student’s t -test was used for A, B, E, and F. The Spearman correlation was used for C and D. * p < 0.05 , ** p < 0.01 . Note: AGA, appropriate for gestational age; As, arsenic; CYR61, cysteine-rich angiogenic inducer 61; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation; SEM, standard error of the mean; SGA, small for gestational age.

    Journal: Environmental Health Perspectives

    Article Title: Effects of Gestational Arsenic Exposures on Placental and Fetal Development in Mice: The Role of Cyr61 m 6 A

    doi: 10.1289/EHP12207

    Figure Lengend Snippet: Based on a case–control study: association among maternal urinary As concentration, plasma CYR61 content, and fetal growth restriction. (A) Maternal urinary As concentration in AGA and SGA infants was determined by hydride generation-atomic fluorescence spectrometry ( n = 45 human sample/group). (B) Maternal plasma CYR61 content in AGA and SGA infants was measured by ELISA ( n = 45 human sample/group). (C) The correlation between maternal urinary As concentration and plasma CYR61 content in AGA infants ( n = 45 human sample). (D) The correlation between maternal urinary As concentration and plasma CYR61 content in SGA infants ( n = 45 human sample). (E) Placental CYR61-positive cells in SGA infants and AGA infants were stained by immunohistochemistry ( n = 18 human sample/group). Representative histology images were showed. Blue arrows indicate CYR61-positive cells. Scale bars, 20 μ m . (F) Placental vimentin, MMP2, and MMP9 were detected by immunoblotting ( n = 18 human sample/group). The data in A and B are expressed as mean ± SD . The data in A–D are represented by dots indicating individual values. The data in E and F are expressed as mean ± SEM . The numeric data are shown in Table S8. Two-tailed Student’s t -test was used for A, B, E, and F. The Spearman correlation was used for C and D. * p < 0.05 , ** p < 0.01 . Note: AGA, appropriate for gestational age; As, arsenic; CYR61, cysteine-rich angiogenic inducer 61; ELISA, enzyme-linked immunosorbent assay; SD, standard deviation; SEM, standard error of the mean; SGA, small for gestational age.

    Article Snippet: The cysteine-rich angiogenic inducer 61 (CYR61) ELISA kit (#CSB-E13884h) was purchased from Cusabio.

    Techniques: Concentration Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry, Western Blot, Two Tailed Test, Standard Deviation

    Primer Sequences for RT-qPCR

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Downregulation of microRNA-224-3p Hampers Retinoblastoma Progression via Activation of the Hippo-YAP Signaling Pathway by Increasing LATS2

    doi: 10.1167/iovs.61.3.32

    Figure Lengend Snippet: Primer Sequences for RT-qPCR

    Article Snippet: The primary antibodies, including LATS2 (1 µg/mL, ab110780), B-cell lymphoma 2 (Bcl-2) (1:1000, ab32124), Bcl-2 associated X protein (Bax) (1:1000, ab3250), vascular endothelial growth factor (VEGF) (1:1000, ab32152), tafazzin (TAZ) (1 µg/mL, ab84927), Yes associated protein (YAP) (1:5000, ab52771), p-YAP (1:10000, ab76252), connective tissue growth factor (CTGF) (1:1000, ab6992), and cysteine rich angiogenic inducer 61 (CYR61) (1 µg/mL, ab24448), were purchased from Abcam, Inc. (Cambridge, UK) except p-TAZ (1:1000, sc-17610-R; Santa Cruz, USA).

    Techniques: Sequencing

    miR-224-3p inhibits the Hippo-YAP signaling pathway by downregulating expression of LATS2. ( A ) Involvement of LATS2 in the Hippo-YAP signaling pathway analyzed on KEGG. ( B ) mRNA expression of TAZ and YAP in cells determined by PT-qPCR. ( C ) Protein expression of TAZ, YAP, p-TAZ, and p-YAP in cells normalized to GAPDH determined by Western blot analysis. ( D ) mRNA expression of TAZ, YAP, CTGF, and CYR61 in cells determined by PT-qPCR. ( E ) Protein expression of TAZ, YAP, CTGF, CYR61, p-TAZ, and p-YAP in cells normalized to GAPDH determined by Western blot analysis. * P < 0.05 versus Y79 cells transfected with NC mimic or oe-NC; # P < 0.05 versus Y79 cells transfected with NC inhibitor or cotransfected with miR-224-3p mimic and oe-NC. Data expressed by mean ± standard deviation among multiple groups were analyzed using 1-way ANOVA, followed by Tukey's post hoc test. The experiment was repeated three times independently.

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Downregulation of microRNA-224-3p Hampers Retinoblastoma Progression via Activation of the Hippo-YAP Signaling Pathway by Increasing LATS2

    doi: 10.1167/iovs.61.3.32

    Figure Lengend Snippet: miR-224-3p inhibits the Hippo-YAP signaling pathway by downregulating expression of LATS2. ( A ) Involvement of LATS2 in the Hippo-YAP signaling pathway analyzed on KEGG. ( B ) mRNA expression of TAZ and YAP in cells determined by PT-qPCR. ( C ) Protein expression of TAZ, YAP, p-TAZ, and p-YAP in cells normalized to GAPDH determined by Western blot analysis. ( D ) mRNA expression of TAZ, YAP, CTGF, and CYR61 in cells determined by PT-qPCR. ( E ) Protein expression of TAZ, YAP, CTGF, CYR61, p-TAZ, and p-YAP in cells normalized to GAPDH determined by Western blot analysis. * P < 0.05 versus Y79 cells transfected with NC mimic or oe-NC; # P < 0.05 versus Y79 cells transfected with NC inhibitor or cotransfected with miR-224-3p mimic and oe-NC. Data expressed by mean ± standard deviation among multiple groups were analyzed using 1-way ANOVA, followed by Tukey's post hoc test. The experiment was repeated three times independently.

    Article Snippet: The primary antibodies, including LATS2 (1 µg/mL, ab110780), B-cell lymphoma 2 (Bcl-2) (1:1000, ab32124), Bcl-2 associated X protein (Bax) (1:1000, ab3250), vascular endothelial growth factor (VEGF) (1:1000, ab32152), tafazzin (TAZ) (1 µg/mL, ab84927), Yes associated protein (YAP) (1:5000, ab52771), p-YAP (1:10000, ab76252), connective tissue growth factor (CTGF) (1:1000, ab6992), and cysteine rich angiogenic inducer 61 (CYR61) (1 µg/mL, ab24448), were purchased from Abcam, Inc. (Cambridge, UK) except p-TAZ (1:1000, sc-17610-R; Santa Cruz, USA).

    Techniques: Expressing, Western Blot, Transfection, Standard Deviation

    A mechanism map depicting the role of the miR-224-3p/LATS2/Hippo-YAP axis in the progression of retinoblastoma. miR-224-3p targets and negatively regulates LATS2 to inhibit the YAP/TAZ phosphorylation, whereby the Hippo signaling pathway activation is inhibited; following that, the downstream genes CTGF and CYR61 are upregulated, by which the apoptosis of retinoblastoma cells is suppressed while proliferation and angiogenesis are promoted.

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Downregulation of microRNA-224-3p Hampers Retinoblastoma Progression via Activation of the Hippo-YAP Signaling Pathway by Increasing LATS2

    doi: 10.1167/iovs.61.3.32

    Figure Lengend Snippet: A mechanism map depicting the role of the miR-224-3p/LATS2/Hippo-YAP axis in the progression of retinoblastoma. miR-224-3p targets and negatively regulates LATS2 to inhibit the YAP/TAZ phosphorylation, whereby the Hippo signaling pathway activation is inhibited; following that, the downstream genes CTGF and CYR61 are upregulated, by which the apoptosis of retinoblastoma cells is suppressed while proliferation and angiogenesis are promoted.

    Article Snippet: The primary antibodies, including LATS2 (1 µg/mL, ab110780), B-cell lymphoma 2 (Bcl-2) (1:1000, ab32124), Bcl-2 associated X protein (Bax) (1:1000, ab3250), vascular endothelial growth factor (VEGF) (1:1000, ab32152), tafazzin (TAZ) (1 µg/mL, ab84927), Yes associated protein (YAP) (1:5000, ab52771), p-YAP (1:10000, ab76252), connective tissue growth factor (CTGF) (1:1000, ab6992), and cysteine rich angiogenic inducer 61 (CYR61) (1 µg/mL, ab24448), were purchased from Abcam, Inc. (Cambridge, UK) except p-TAZ (1:1000, sc-17610-R; Santa Cruz, USA).

    Techniques: Activation Assay