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Anhydrous, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Title: Complete Kinetic Characterization of Enzyme Inhibition in a Single Isothermal Titration Calorimetric Experiment
Article Snippet: Techniques for rapidly measuring both the strength and mode of enzyme inhibitors are crucial to lead generation and optimization in drug development.. Isothermal titration calorimetry (ITC) is emerging as a powerful tool for measuring enzyme kinetics with distinct advantages over traditional techniques.. ITC measures heat flow, a feature of nearly all chemical reactions, and gives an instantaneous read-out of enzyme velocity, eliminating the need for artificial substrates or post-reaction processing.

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Screening of the candidate genes based on MeRIP-seq and RNA-seq. ( A ) The proportion analysis for specific m 6 A binding sites in MeRIP-seq in three groups. ( B ) Top sequence motif identified from MeRIP-seq peaks in three groups. ( C ) Peak distribution of m 6 A modification in meRIP-seq results between the two compared groups. ( D ) Volcano plot of MeRIP-Seq showed the changed m 6 A peaks between two compared groups. Genes with a 2-fold differentially m 6 A-modified genes and FDR ≤ 0.01 were highlighted in red for hyper-methylation and blue for hypo-methylation. ( E ) KEGG pathway enrichment analysis of MeRIP-seq data. ( F ) Expression heatmap of all genes in the dataset under fatty acid metabolism pathway. Heatmap of the data from RNA-seq. Genes with significant changes in expression ( Eci1 , Ggct and Mlycd ) were labeled in red. ( G ) MeRIP-qPCR assay confirmed the results from MeRIP-seq ( n = 4 per group). ( H ) Eci1 mRNA expression was performed using qRT-PCR ( n = 6 per group). ( I ) RIP assay was performed using IgG or FTO antibody. The enrichment of Eci1 mRNA was measured by qRT-PCR assay ( n = 3 per group). ( J ) Eci1 protein expression was performed using Western blot. ( K ) Fold change in Eci1 protein expression ( n = 3 per group). ( L ) FFA levels and TG content in plasma ( n = 7 per group). ( M ) FFA levels and TG content in myocardial tissues ( n = 7 per group). FFA, free fatty acid; TG, triglyceride. Statistical significance is denoted by: ** p < 0.01, *** p < 0.001.

Journal: Biomolecules

Article Title: FTO-Eci1 Axis Mediates Exercise-Induced Cardioprotection in Pressure Overload Mice

doi: 10.3390/biom16010098

Figure Lengend Snippet: Screening of the candidate genes based on MeRIP-seq and RNA-seq. ( A ) The proportion analysis for specific m 6 A binding sites in MeRIP-seq in three groups. ( B ) Top sequence motif identified from MeRIP-seq peaks in three groups. ( C ) Peak distribution of m 6 A modification in meRIP-seq results between the two compared groups. ( D ) Volcano plot of MeRIP-Seq showed the changed m 6 A peaks between two compared groups. Genes with a 2-fold differentially m 6 A-modified genes and FDR ≤ 0.01 were highlighted in red for hyper-methylation and blue for hypo-methylation. ( E ) KEGG pathway enrichment analysis of MeRIP-seq data. ( F ) Expression heatmap of all genes in the dataset under fatty acid metabolism pathway. Heatmap of the data from RNA-seq. Genes with significant changes in expression ( Eci1 , Ggct and Mlycd ) were labeled in red. ( G ) MeRIP-qPCR assay confirmed the results from MeRIP-seq ( n = 4 per group). ( H ) Eci1 mRNA expression was performed using qRT-PCR ( n = 6 per group). ( I ) RIP assay was performed using IgG or FTO antibody. The enrichment of Eci1 mRNA was measured by qRT-PCR assay ( n = 3 per group). ( J ) Eci1 protein expression was performed using Western blot. ( K ) Fold change in Eci1 protein expression ( n = 3 per group). ( L ) FFA levels and TG content in plasma ( n = 7 per group). ( M ) FFA levels and TG content in myocardial tissues ( n = 7 per group). FFA, free fatty acid; TG, triglyceride. Statistical significance is denoted by: ** p < 0.01, *** p < 0.001.

Article Snippet: Eci1-LoxP-targeted (Eci1 fl/fl ) mice (Name: C57BL/6J-Eci1 em1(flox)Cya , Serial Number: CKOCMP-13177-Eci1-B6J-VA) were created by Cyagen Biosciences (Suzhou, China).

Techniques: RNA Sequencing, Binding Assay, Sequencing, Modification, Methylation, Expressing, Labeling, Quantitative RT-PCR, Western Blot, Clinical Proteomics

$The role of Eci1 in ameliorating heart failure by exercise intervention. ( A ) Schematic diagram of exercise intervention. Arrows indicate timing of tamoxifen administration. ( B ) Verification of Eci1 knockdown efficiency in myocardial tissues ( n = 3 per group). ( C ) Cardiac function was evaluated by calculating EF and FS ( n = 8 per group). EF, ejection fraction; FS, fractional shortening. ( D ) Representative images of Masson’s trichrome staining and H&E staining of heart tissue. Quantitative results of Masson’s trichrome staining ( n = 8 per group). Scale bar, 200 µm. ( E ) Wheat germ agglutinin (WGA, green) staining showed cardiomyocyte membrane staining and quantification results for the cardiomyocyte cross sectional area ( n = 8 per group). Scale bar, 100 µm. ( F ) Oil Red O staining and lipid content quantification ( n = 4 per group). Scale bar, 100 µm. ( G ) FFA levels in plasma and myocardial tissues ( n = 7 per group). FFA, free fatty acid. ( H ) Apoptosis related proteins were detected by Western blot ( n = 3 per group). ( I ) Relative ATP content of myocardial tissues (normalized using protein concentration, n = 6 per group). * p < 0.05, ** p < 0.01, *** p < 0.001, ns indicates p > 0.05.$

Journal: Biomolecules

Article Title: FTO-Eci1 Axis Mediates Exercise-Induced Cardioprotection in Pressure Overload Mice

doi: 10.3390/biom16010098

Figure Lengend Snippet: The role of Eci1 in ameliorating heart failure by exercise intervention. ( A ) Schematic diagram of exercise intervention. Arrows indicate timing of tamoxifen administration. ( B ) Verification of Eci1 knockdown efficiency in myocardial tissues ( n = 3 per group). ( C ) Cardiac function was evaluated by calculating EF and FS ( n = 8 per group). EF, ejection fraction; FS, fractional shortening. ( D ) Representative images of Masson’s trichrome staining and H&E staining of heart tissue. Quantitative results of Masson’s trichrome staining ( n = 8 per group). Scale bar, 200 µm. ( E ) Wheat germ agglutinin (WGA, green) staining showed cardiomyocyte membrane staining and quantification results for the cardiomyocyte cross sectional area ( n = 8 per group). Scale bar, 100 µm. ( F ) Oil Red O staining and lipid content quantification ( n = 4 per group). Scale bar, 100 µm. ( G ) FFA levels in plasma and myocardial tissues ( n = 7 per group). FFA, free fatty acid. ( H ) Apoptosis related proteins were detected by Western blot ( n = 3 per group). ( I ) Relative ATP content of myocardial tissues (normalized using protein concentration, n = 6 per group). * p < 0.05, ** p < 0.01, *** p < 0.001, ns indicates p > 0.05.

Article Snippet: Eci1-LoxP-targeted (Eci1 fl/fl ) mice (Name: C57BL/6J-Eci1 em1(flox)Cya , Serial Number: CKOCMP-13177-Eci1-B6J-VA) were created by Cyagen Biosciences (Suzhou, China).

Techniques: Knockdown, Staining, Membrane, Clinical Proteomics, Western Blot, Protein Concentration

The role of Eci1 in cardiomyocyte hypertrophy induced by Ang II. ( A ) Detection of cell apoptosis by flow cytometry analysis. ( B ) The statistic results of cell apoptosis by flow cytometry ( n = 3 per group). ( C ) The apoptosis related proteins were detected ( n = 3 per group). ( D ) Representative image of Oil Red O stain. ( E ) Evaluation of lipid accumulation by Oil Red O staining ( n = 3 per group). Scale bar, 50 µm. ( F ) FFA levels and TG content in each group ( n = 3 per group). FFA, free fatty acid; TG, triglyceride. ( G ) Relative ATP content of cardiomyocytes (normalized using protein concentration, n = 3 per group). Statistical significance is denoted by: * p < 0.05, ** p < 0.01, *** p < 0.001, ns indicates p > 0.05. OE-NC, overexpression-control. OE-Eci1, overexpression-Eci1. shNC, shRNA-control. shEci1, shRNA-Eci1.

Journal: Biomolecules

Article Title: FTO-Eci1 Axis Mediates Exercise-Induced Cardioprotection in Pressure Overload Mice

doi: 10.3390/biom16010098

Figure Lengend Snippet: The role of Eci1 in cardiomyocyte hypertrophy induced by Ang II. ( A ) Detection of cell apoptosis by flow cytometry analysis. ( B ) The statistic results of cell apoptosis by flow cytometry ( n = 3 per group). ( C ) The apoptosis related proteins were detected ( n = 3 per group). ( D ) Representative image of Oil Red O stain. ( E ) Evaluation of lipid accumulation by Oil Red O staining ( n = 3 per group). Scale bar, 50 µm. ( F ) FFA levels and TG content in each group ( n = 3 per group). FFA, free fatty acid; TG, triglyceride. ( G ) Relative ATP content of cardiomyocytes (normalized using protein concentration, n = 3 per group). Statistical significance is denoted by: * p < 0.05, ** p < 0.01, *** p < 0.001, ns indicates p > 0.05. OE-NC, overexpression-control. OE-Eci1, overexpression-Eci1. shNC, shRNA-control. shEci1, shRNA-Eci1.

Article Snippet: Eci1-LoxP-targeted (Eci1 fl/fl ) mice (Name: C57BL/6J-Eci1 em1(flox)Cya , Serial Number: CKOCMP-13177-Eci1-B6J-VA) were created by Cyagen Biosciences (Suzhou, China).

Techniques: Flow Cytometry, Staining, Protein Concentration, Over Expression, Control, shRNA

FTO promoted the expression of Eci1 dependent on its m 6 A demethylase activity. ( A ) Flow cytometry analysis was used to detect cell apoptosis ( n = 3 per group). ( B ) Apoptosis related proteins were detected by Western blot ( n = 3 per group). ( C ) Eci1 protein expression was demonstrated by Western blot ( n = 3 per group). ( D ) The expression of Eci1 was measured by qRT-PCR. ( E ) Total m 6 A content was detected. ( F ) Oil Red O staining and lipid content quantification ( n = 3 per group). Scale bar, 50 µm. ( G ) FFA levels and TG content in each group ( n = 3 per group). FFA, free fatty acid; TG, triglyceride. ( H ) Diagram of the proposed molecular mechanism of the cardioprotective effect of exercise intervention. The downward arrow denotes a decrease, whereas the upward arrow denotes an increase in content, production, or expression. The diagram was drawn by using Figdraw ( www.figdraw.com ). Statistical significance is denoted by: * p < 0.05, ** p < 0.01, *** p < 0.001, ns indicates p > 0.05. shNC, shRNA-control. shEci1, shRNA-FTO. OE-NC, overexpression-control. OE-FTO, overexpression-FTO. OE-FTO (MUT), overexpression-FTO (mutated FTO, R96Q).

Journal: Biomolecules

Article Title: FTO-Eci1 Axis Mediates Exercise-Induced Cardioprotection in Pressure Overload Mice

doi: 10.3390/biom16010098

Figure Lengend Snippet: FTO promoted the expression of Eci1 dependent on its m 6 A demethylase activity. ( A ) Flow cytometry analysis was used to detect cell apoptosis ( n = 3 per group). ( B ) Apoptosis related proteins were detected by Western blot ( n = 3 per group). ( C ) Eci1 protein expression was demonstrated by Western blot ( n = 3 per group). ( D ) The expression of Eci1 was measured by qRT-PCR. ( E ) Total m 6 A content was detected. ( F ) Oil Red O staining and lipid content quantification ( n = 3 per group). Scale bar, 50 µm. ( G ) FFA levels and TG content in each group ( n = 3 per group). FFA, free fatty acid; TG, triglyceride. ( H ) Diagram of the proposed molecular mechanism of the cardioprotective effect of exercise intervention. The downward arrow denotes a decrease, whereas the upward arrow denotes an increase in content, production, or expression. The diagram was drawn by using Figdraw ( www.figdraw.com ). Statistical significance is denoted by: * p < 0.05, ** p < 0.01, *** p < 0.001, ns indicates p > 0.05. shNC, shRNA-control. shEci1, shRNA-FTO. OE-NC, overexpression-control. OE-FTO, overexpression-FTO. OE-FTO (MUT), overexpression-FTO (mutated FTO, R96Q).

Article Snippet: Eci1-LoxP-targeted (Eci1 fl/fl ) mice (Name: C57BL/6J-Eci1 em1(flox)Cya , Serial Number: CKOCMP-13177-Eci1-B6J-VA) were created by Cyagen Biosciences (Suzhou, China).

Techniques: Expressing, Activity Assay, Flow Cytometry, Western Blot, Quantitative RT-PCR, Staining, shRNA, Control, Over Expression

Bacterial growth in different strains of Proteus mirabilis . Growth curves of P. mirabilis C02011, B02005, and P. mirabilis American Type Culture Collection (ATCC) 29906 detected by Densimat photometer.

Journal: Frontiers in Microbiology

Article Title: Characterization of a Novel Diarrheagenic Strain of Proteus mirabilis Associated With Food Poisoning in China

doi: 10.3389/fmicb.2019.02810

Figure Lengend Snippet: Bacterial growth in different strains of Proteus mirabilis . Growth curves of P. mirabilis C02011, B02005, and P. mirabilis American Type Culture Collection (ATCC) 29906 detected by Densimat photometer.

Article Snippet: In the results, P. mirabilis B02005 strain (11.70 ± 0.36 × 10 4 CFU/well) is more adhesive than P. mirabilis C02011 strain (7.90 ± 0.26 × 10 4 CFU/well), and P. mirabilis ATCC 29906 strain (3.63 ± 0.32 × 10 4 CFU/well): P. mirabilis B02005 > P. mirabilis C02011 > P. mirabilis ATCC 29906 [ P ( B 02005 vs. C02011) = 0.000, P ( B 02005 vs. P. mirabilis ATCC 29906 ) = 0.000, ∗∗ P < 0.01] ( ).

Techniques:

HT-29, LoVo, and Caco-2 cells were infected with Proteus mirabilis C02011. (A) The adhesion ability of P. mirabilis C02011 to the three kinds of cells (HT-29, Caco-2, and LoVo) was examined. (B) The invasion ability of P. mirabilis C02011 to the three kinds of cells (HT-29, Caco-2, and LoVo) were examined. (C–E) Giemsa staining of HT-29, LoVo, and Caco-2 cells infected with P. mirabilis C02011. (C) Giemsa staining of HT-29 cells infected with P. mirabilis C02011. (D) Giemsa staining of LoVo cells infected with P. mirabilis C02011. (E) Giemsa staining of Caco-2 cells infected with P. mirabilis C02011. ∗ P < 0.05, ∗∗ P < 0.01; NS, no significance.

Journal: Frontiers in Microbiology

Article Title: Characterization of a Novel Diarrheagenic Strain of Proteus mirabilis Associated With Food Poisoning in China

doi: 10.3389/fmicb.2019.02810

Figure Lengend Snippet: HT-29, LoVo, and Caco-2 cells were infected with Proteus mirabilis C02011. (A) The adhesion ability of P. mirabilis C02011 to the three kinds of cells (HT-29, Caco-2, and LoVo) was examined. (B) The invasion ability of P. mirabilis C02011 to the three kinds of cells (HT-29, Caco-2, and LoVo) were examined. (C–E) Giemsa staining of HT-29, LoVo, and Caco-2 cells infected with P. mirabilis C02011. (C) Giemsa staining of HT-29 cells infected with P. mirabilis C02011. (D) Giemsa staining of LoVo cells infected with P. mirabilis C02011. (E) Giemsa staining of Caco-2 cells infected with P. mirabilis C02011. ∗ P < 0.05, ∗∗ P < 0.01; NS, no significance.

Techniques: Infection, Staining

Adhesion and invasion of Proteus mirabilis C02011 strain, P. mirabilis B02005, and P. mirabilis American Type Culture Collection (ATCC) 29906 strains to Caco-2 cells. (A) Caco-2 cells were infected with P. mirabilis C02011, P. mirabilis B02005, and P. mirabilis ATCC 29906 strains for 2 h. The adhesion abilities of different strains were recorded. (B) Caco-2 cells were infected with P. mirabilis C02011, P. mirabilis B02005, and P. mirabilis ATCC 29906 for 2 h, followed by treatment with gentamicin (200 μg/ml) for 2 h. The invasion abilities of different strains were recorded. (C–E) Transmission electron microscopy (TEM) images were collected for observation. (C) Morphological observation of P. mirabilis C02011 (×15,000 magnification). (D) Morphological observation of uninfected Caco-2 cell (×7,000 magnification). (E) Morphological observation of Caco-2 cell infected with P. mirabilis C02011 (×7,000 magnification). ∗∗ P < 0.01.

Journal: Frontiers in Microbiology

Article Title: Characterization of a Novel Diarrheagenic Strain of Proteus mirabilis Associated With Food Poisoning in China

doi: 10.3389/fmicb.2019.02810

Figure Lengend Snippet: Adhesion and invasion of Proteus mirabilis C02011 strain, P. mirabilis B02005, and P. mirabilis American Type Culture Collection (ATCC) 29906 strains to Caco-2 cells. (A) Caco-2 cells were infected with P. mirabilis C02011, P. mirabilis B02005, and P. mirabilis ATCC 29906 strains for 2 h. The adhesion abilities of different strains were recorded. (B) Caco-2 cells were infected with P. mirabilis C02011, P. mirabilis B02005, and P. mirabilis ATCC 29906 for 2 h, followed by treatment with gentamicin (200 μg/ml) for 2 h. The invasion abilities of different strains were recorded. (C–E) Transmission electron microscopy (TEM) images were collected for observation. (C) Morphological observation of P. mirabilis C02011 (×15,000 magnification). (D) Morphological observation of uninfected Caco-2 cell (×7,000 magnification). (E) Morphological observation of Caco-2 cell infected with P. mirabilis C02011 (×7,000 magnification). ∗∗ P < 0.01.

Techniques: Infection, Transmission Assay, Electron Microscopy

Diarrhea induced by Proteus mirabilis C02011 was observed in BALB/c mice. (A) Streptomycin-treated mice were gavaged with P. mirabilis C02011, P. mirabilis B02005, and normal saline. Clinical symptoms and colonic lesions were evaluated by indicators. (B) Feces images of mice in different groups. (C) Feces water content of mice in different groups was recorded at 0, 1, 3, and 5 h after infection. (D) Colon images in different groups. (E) Anal region images of mice in different groups. (F) Mouse colon length was measured in different groups. (G) The level of TNF-α in colon tissues. ∗ P < 0.05, ∗∗ P < 0.01.

Journal: Frontiers in Microbiology

Article Title: Characterization of a Novel Diarrheagenic Strain of Proteus mirabilis Associated With Food Poisoning in China

doi: 10.3389/fmicb.2019.02810

Figure Lengend Snippet: Diarrhea induced by Proteus mirabilis C02011 was observed in BALB/c mice. (A) Streptomycin-treated mice were gavaged with P. mirabilis C02011, P. mirabilis B02005, and normal saline. Clinical symptoms and colonic lesions were evaluated by indicators. (B) Feces images of mice in different groups. (C) Feces water content of mice in different groups was recorded at 0, 1, 3, and 5 h after infection. (D) Colon images in different groups. (E) Anal region images of mice in different groups. (F) Mouse colon length was measured in different groups. (G) The level of TNF-α in colon tissues. ∗ P < 0.05, ∗∗ P < 0.01.

Techniques: Saline, Infection

Hematoxylin and eosin (H&E) staining of the small intestine and large intestine tissues in Proteus mirabilis C02011, B02005, and control groups. (A) Histopathological observations of small bowel in P. mirabilis C02011, B02005, and control groups (×100, ×200, and ×400 magnification). (B) Histopathological observations of large bowel in P. mirabilis C02011, B02005, and control groups (×100, ×200, and ×400 magnification). (C) The average for each element in the histological score (including inflammation, extent, regeneration, crypt damage, and percent involvement) for mice in different groups. ∗∗ P < 0.01.

Journal: Frontiers in Microbiology

Article Title: Characterization of a Novel Diarrheagenic Strain of Proteus mirabilis Associated With Food Poisoning in China

doi: 10.3389/fmicb.2019.02810

Figure Lengend Snippet: Hematoxylin and eosin (H&E) staining of the small intestine and large intestine tissues in Proteus mirabilis C02011, B02005, and control groups. (A) Histopathological observations of small bowel in P. mirabilis C02011, B02005, and control groups (×100, ×200, and ×400 magnification). (B) Histopathological observations of large bowel in P. mirabilis C02011, B02005, and control groups (×100, ×200, and ×400 magnification). (C) The average for each element in the histological score (including inflammation, extent, regeneration, crypt damage, and percent involvement) for mice in different groups. ∗∗ P < 0.01.

Techniques: Staining, Control

The expressions of protein (occludin) in colon tissues induced by Proteus mirabilis . (A) The expressions of protein (occludin) detected by western blotting (WB). (B) The occludin/β-actin intensity ratio in different groups. (C) The expressions of protein (occludin) in different groups detected by immunohistochemistry (IHC). (D–F) Large intestinal sections were stained by IHC (occludin) antibody in different groups: (D) IHC image in C02011 group; (E) IHC image in B02005 group; (F) IHC image in control group. ∗ P < 0.05, ∗∗ P < 0.01; NS, no significance.

Journal: Frontiers in Microbiology

Article Title: Characterization of a Novel Diarrheagenic Strain of Proteus mirabilis Associated With Food Poisoning in China

doi: 10.3389/fmicb.2019.02810

Figure Lengend Snippet: The expressions of protein (occludin) in colon tissues induced by Proteus mirabilis . (A) The expressions of protein (occludin) detected by western blotting (WB). (B) The occludin/β-actin intensity ratio in different groups. (C) The expressions of protein (occludin) in different groups detected by immunohistochemistry (IHC). (D–F) Large intestinal sections were stained by IHC (occludin) antibody in different groups: (D) IHC image in C02011 group; (E) IHC image in B02005 group; (F) IHC image in control group. ∗ P < 0.05, ∗∗ P < 0.01; NS, no significance.

Techniques: Western Blot, Immunohistochemistry, Staining, Control

Adhesion and invasion of Proteus mirabilis C02011 strain and its mutant strain to Caco-2 cells. (A) Bacterial growth of P. mirabilis C02011 and its mutant strain detected by Densimat photometer. (B) Caco-2 cells were infected with P. mirabilis C02011 and its mutant strain for 2 h. The adhesion abilities of different strains were recorded. (C) Caco-2 cells were infected with P. mirabilis C02011 and its mutant strain for 2 h, followed by treatment with gentamicin (200 μg/ml) for 2 h. The invasion abilities of different strains were recorded. ∗ P < 0.05; NS, no significance.

Journal: Frontiers in Microbiology

Article Title: Characterization of a Novel Diarrheagenic Strain of Proteus mirabilis Associated With Food Poisoning in China

doi: 10.3389/fmicb.2019.02810

Figure Lengend Snippet: Adhesion and invasion of Proteus mirabilis C02011 strain and its mutant strain to Caco-2 cells. (A) Bacterial growth of P. mirabilis C02011 and its mutant strain detected by Densimat photometer. (B) Caco-2 cells were infected with P. mirabilis C02011 and its mutant strain for 2 h. The adhesion abilities of different strains were recorded. (C) Caco-2 cells were infected with P. mirabilis C02011 and its mutant strain for 2 h, followed by treatment with gentamicin (200 μg/ml) for 2 h. The invasion abilities of different strains were recorded. ∗ P < 0.05; NS, no significance.

Techniques: Mutagenesis, Infection

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