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  • 99
    Millipore prostaglandin e2
    Prostaglandin E2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1618 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/prostaglandin e2/product/Millipore
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    92
    Cayman Chemical prostaglandin e2
    Model for <t>PGE2</t> signaling to expand MuSC function in regeneration. Schematic of the role of PGE2 in MuSCs. After injury, PGE2 released into the muscle niche acts on the EP4 receptor, which signals through cAMP/phospho-CREB leading to the expression of
    Prostaglandin E2, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 92/100, based on 1131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore e2
    Model for <t>PGE2</t> signaling to expand MuSC function in regeneration. Schematic of the role of PGE2 in MuSCs. After injury, PGE2 released into the muscle niche acts on the EP4 receptor, which signals through cAMP/phospho-CREB leading to the expression of
    E2, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 2157 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 17β estradiol e2
    Model for <t>PGE2</t> signaling to expand MuSC function in regeneration. Schematic of the role of PGE2 in MuSCs. After injury, PGE2 released into the muscle niche acts on the EP4 receptor, which signals through cAMP/phospho-CREB leading to the expression of
    17β Estradiol E2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 983 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cayman Chemical pge2 eia kit
    Model for <t>PGE2</t> signaling to expand MuSC function in regeneration. Schematic of the role of PGE2 in MuSCs. After injury, PGE2 released into the muscle niche acts on the EP4 receptor, which signals through cAMP/phospho-CREB leading to the expression of
    Pge2 Eia Kit, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 99/100, based on 878 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Tocris prostaglandin e2
    NK-92MI cell migration and subsequent cytotoxicity toward A549/GFP spheroids. NK-92MI (NK) cell migration (blue) was evaluated in the presence and absence of stromal-cell derived factor-1 (SDF-1α) and/or <t>prostaglandin</t> E2 (PGE) in the medium. Percent migration (left axis) was calculated as a ratio of cells in basolateral chamber compared with total cells seeded in insert. Cytotoxicity of A549/GFP cells (green) was evaluated after exposure to NK-92MI (NK) cells in the presence and absence of SDF-1α and/or PGE in the medium. Percent viability (right axis) was calculated via flow cytometry by enumerating GFP-positive A549 cells relative to spheroids without effector cells. Data are shown as the average of two independent studies, n = 24 wells. * p
    Prostaglandin E2, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pge2  (Abcam)
    95
    Abcam pge2
    Multivariate data analyses of factors responsible for vascular dysfunction. (A) Scores scatter plot t1 vs. t2 resulting after applying PCA to endometrial expression of various angiogenic and vasoactive factors of controls (green) and IRSM (blue), (B) scores scatter plot t1 vs t2 resulting after applying PLS-DA to endometrial expression of various angiogenic and vasoactive factors of controls (green) and IRSM (blue), (C) Results from the permutation test for the IRSM group suggest a valid PLS-DA model. The vertical axis is the R2 and Q2 values of each model and the horizontal axis shows the correlation between the permuted class vectors and the original class vector. The original class has the correlation 1.0 with itself, defining the high point on the horizontal axis. All R2 and Q2 values calculated from the permuted data are lower than the original model in the validation plot. Y-axis intercepts: R2= (0.0, 0.0189), Q2= (0.0, −0.123). (D) Loading scatter plot indicates factors IL-1β, TNF-α, IFN-γ, TGF-β1, <t>PGE2</t> are upregulated in IRSM and factors IL-2, IL-4, IL-6, IL-8, IL-10, VEGF, ADM, eNOS, NO are upregulated in controls.
    Pge2, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 216 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Cell Signaling Technology Inc cyclin e2
    Ectopic Expression of miR-411 Induced Cell Cycles Arrest in G2/M Phase by Up-regulating p21 Protein in Human BC Cells (A) Cell lysates from the indicated cells were subjected to western blot to analyze expression of p21, <t>cyclin</t> B1, cyclin A2, and cyclin E2. β-actin was used as an internal control. (B) Western blot was used to determine the expression of p21 in T24 (miR-411/HA-MLLT11) and UMUC3 (miR-411/HA-MLLT11) cells in comparison to their vector control transfectants. β-actin was used as an internal control. (C) UMUC3 (miR-411) cells stably transfected with shRNA targeting human p21 or its nonsense control plasmid. The cell lysates from the indicated transfectants were subjected to western blot to assess the knockdown efficiency of the p21 protein. (D) UMUC3 (miR-411/nonsense), UMUC3 (miR-411/shp21 No. 3), and UMUC3 (miR-411/shp21 No. 6) cells were subjected to soft-agar assay to determine the effect of p21 on miR-411-induced inhibition of anchorage-independent growth. The images of colonies from the indicated cells were captured under microscopy after 3 weeks of incubation. (E) The number of colonies was scored and presented as colonies per 10,000 cells. The symbol (*) indicates a significant increase in comparison with UMUC3 (miR-411/Nonsense) transfectants (*p
    Cyclin E2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 425 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore estradiol e2
    Ectopic Expression of miR-411 Induced Cell Cycles Arrest in G2/M Phase by Up-regulating p21 Protein in Human BC Cells (A) Cell lysates from the indicated cells were subjected to western blot to analyze expression of p21, <t>cyclin</t> B1, cyclin A2, and cyclin E2. β-actin was used as an internal control. (B) Western blot was used to determine the expression of p21 in T24 (miR-411/HA-MLLT11) and UMUC3 (miR-411/HA-MLLT11) cells in comparison to their vector control transfectants. β-actin was used as an internal control. (C) UMUC3 (miR-411) cells stably transfected with shRNA targeting human p21 or its nonsense control plasmid. The cell lysates from the indicated transfectants were subjected to western blot to assess the knockdown efficiency of the p21 protein. (D) UMUC3 (miR-411/nonsense), UMUC3 (miR-411/shp21 No. 3), and UMUC3 (miR-411/shp21 No. 6) cells were subjected to soft-agar assay to determine the effect of p21 on miR-411-induced inhibition of anchorage-independent growth. The images of colonies from the indicated cells were captured under microscopy after 3 weeks of incubation. (E) The number of colonies was scored and presented as colonies per 10,000 cells. The symbol (*) indicates a significant increase in comparison with UMUC3 (miR-411/Nonsense) transfectants (*p
    Estradiol E2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 199 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    steraloids inc e2 bsa
    Ectopic Expression of miR-411 Induced Cell Cycles Arrest in G2/M Phase by Up-regulating p21 Protein in Human BC Cells (A) Cell lysates from the indicated cells were subjected to western blot to analyze expression of p21, <t>cyclin</t> B1, cyclin A2, and cyclin E2. β-actin was used as an internal control. (B) Western blot was used to determine the expression of p21 in T24 (miR-411/HA-MLLT11) and UMUC3 (miR-411/HA-MLLT11) cells in comparison to their vector control transfectants. β-actin was used as an internal control. (C) UMUC3 (miR-411) cells stably transfected with shRNA targeting human p21 or its nonsense control plasmid. The cell lysates from the indicated transfectants were subjected to western blot to assess the knockdown efficiency of the p21 protein. (D) UMUC3 (miR-411/nonsense), UMUC3 (miR-411/shp21 No. 3), and UMUC3 (miR-411/shp21 No. 6) cells were subjected to soft-agar assay to determine the effect of p21 on miR-411-induced inhibition of anchorage-independent growth. The images of colonies from the indicated cells were captured under microscopy after 3 weeks of incubation. (E) The number of colonies was scored and presented as colonies per 10,000 cells. The symbol (*) indicates a significant increase in comparison with UMUC3 (miR-411/Nonsense) transfectants (*p
    E2 Bsa, supplied by steraloids inc, used in various techniques. Bioz Stars score: 90/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Xenobiotics pdc e2
    Germline-encoded clone strongly reacted to xenobiotics A) AA sequences of cross-reactive clones S1-8 and S1-9 aligned with IGHV 3-30 germline sequence. B) Schematic representation of conversion of germline IGHV3-30-reverted antibody generation. C) A side-by-side comparison of antigen reactivities against <t>PDC-E2</t> and 2OA-BSA using immunoblots. Equal concentrations of mAbs were used to probe PDC-E2 and 2OA-BSA.
    Pdc E2, supplied by Xenobiotics, used in various techniques. Bioz Stars score: 89/100, based on 134 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cayman Chemical 15 keto pge2
    Germline-encoded clone strongly reacted to xenobiotics A) AA sequences of cross-reactive clones S1-8 and S1-9 aligned with IGHV 3-30 germline sequence. B) Schematic representation of conversion of germline IGHV3-30-reverted antibody generation. C) A side-by-side comparison of antigen reactivities against <t>PDC-E2</t> and 2OA-BSA using immunoblots. Equal concentrations of mAbs were used to probe PDC-E2 and 2OA-BSA.
    15 Keto Pge2, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 93/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Diagnosys LLC espion e2 system
    Germline-encoded clone strongly reacted to xenobiotics A) AA sequences of cross-reactive clones S1-8 and S1-9 aligned with IGHV 3-30 germline sequence. B) Schematic representation of conversion of germline IGHV3-30-reverted antibody generation. C) A side-by-side comparison of antigen reactivities against <t>PDC-E2</t> and 2OA-BSA using immunoblots. Equal concentrations of mAbs were used to probe PDC-E2 and 2OA-BSA.
    Espion E2 System, supplied by Diagnosys LLC, used in various techniques. Bioz Stars score: 89/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Model for PGE2 signaling to expand MuSC function in regeneration. Schematic of the role of PGE2 in MuSCs. After injury, PGE2 released into the muscle niche acts on the EP4 receptor, which signals through cAMP/phospho-CREB leading to the expression of

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: Model for PGE2 signaling to expand MuSC function in regeneration. Schematic of the role of PGE2 in MuSCs. After injury, PGE2 released into the muscle niche acts on the EP4 receptor, which signals through cAMP/phospho-CREB leading to the expression of

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Expressing

    PGE2 promotes MuSC proliferation. ( A ) PGE2 levels assayed by ELISA after cryoinjury for TA ( n = 3 mice per condition). Control refers to the contralateral uninjured leg. ( B ) Proliferation measured by the metabolic viability assay VisionBlue after treatment

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: PGE2 promotes MuSC proliferation. ( A ) PGE2 levels assayed by ELISA after cryoinjury for TA ( n = 3 mice per condition). Control refers to the contralateral uninjured leg. ( B ) Proliferation measured by the metabolic viability assay VisionBlue after treatment

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Enzyme-linked Immunosorbent Assay, Mouse Assay, Viability Assay

    Muscle mass is not altered after PGE2 loss of function after injury. ( A ) Mass of TAs of control or MuSC-specific EP4 conditional knockout mice at day 14 after injury. ( B ) Mass of vehicle or NSAID-treated TAs at day 14 after injury. Mann–Whitney

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: Muscle mass is not altered after PGE2 loss of function after injury. ( A ) Mass of TAs of control or MuSC-specific EP4 conditional knockout mice at day 14 after injury. ( B ) Mass of vehicle or NSAID-treated TAs at day 14 after injury. Mann–Whitney

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Knock-Out, Mouse Assay, MANN-WHITNEY

    PGE2 direct injection augments muscle regeneration without promoting hypertrophy. ( A ) Representative cross-sectional images of the TA showing a GFP + MuSC after engraftment of freshly sorted GFP/luc-labeled MuSCs (250 cells) coinjected with PGE2 at 8 wk

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: PGE2 direct injection augments muscle regeneration without promoting hypertrophy. ( A ) Representative cross-sectional images of the TA showing a GFP + MuSC after engraftment of freshly sorted GFP/luc-labeled MuSCs (250 cells) coinjected with PGE2 at 8 wk

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Injection, Labeling

    PGE2 treatment augments muscle regeneration. ( A ) Engraftment of freshly sorted GFP/luc-labeled MuSCs (250 cells) coinjected with vehicle or PGE2. ( Top ) Transplant scheme. ( Bottom ) BLI signals after transplant expressed as average radiance (p·s

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: PGE2 treatment augments muscle regeneration. ( A ) Engraftment of freshly sorted GFP/luc-labeled MuSCs (250 cells) coinjected with vehicle or PGE2. ( Top ) Transplant scheme. ( Bottom ) BLI signals after transplant expressed as average radiance (p·s

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Labeling

    Transcriptome analysis of PGE2-treated MuSCs. ( A ) Heat map of the transcriptome of vehicle or PGE2-treated MuSCs after 24 h shown as expression fold-change over vehicle-treated MuSCs. ( B ) Enriched molecular and cellular function pathways of the differentially

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: Transcriptome analysis of PGE2-treated MuSCs. ( A ) Heat map of the transcriptome of vehicle or PGE2-treated MuSCs after 24 h shown as expression fold-change over vehicle-treated MuSCs. ( B ) Enriched molecular and cellular function pathways of the differentially

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Expressing, Cell Function Assay

    Transient increase in PGE2 in damaged muscle tissues accelerates MuSC proliferation. ( A ) Expression of Ptger4 in freshly isolated MuSCs from uninjured mouse hindlimbs (Fresh MuSCs), MuSCs cultured for 2 d on hydrogels (Cultured MuSCs), primary myoblasts

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: Transient increase in PGE2 in damaged muscle tissues accelerates MuSC proliferation. ( A ) Expression of Ptger4 in freshly isolated MuSCs from uninjured mouse hindlimbs (Fresh MuSCs), MuSCs cultured for 2 d on hydrogels (Cultured MuSCs), primary myoblasts

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Expressing, Isolation, Cell Culture

    Nurr1 is a downstream effector of PGE2/EP4 signaling in MuSCs. ( A ) Heat map of differentially expressed transcription factors in vehicle or PGE2-treated MuSCs after 24 h. ( B ) Expression of Nurr1 after TA muscle injury (notexin) ( n = 3 mice per timepoint).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: Nurr1 is a downstream effector of PGE2/EP4 signaling in MuSCs. ( A ) Heat map of differentially expressed transcription factors in vehicle or PGE2-treated MuSCs after 24 h. ( B ) Expression of Nurr1 after TA muscle injury (notexin) ( n = 3 mice per timepoint).

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Expressing, Mouse Assay

    Loss of function of PGE2 signaling in MuSCs impairs muscle regeneration and strength. ( A–H ) TAs of Pax7-specific EP4 conditional knockout mice ( Pax7 CreERT2 ;EP4 f/f , EP4 cKO) treated with tamoxifen (TAM) were assayed at 7 ( C and E ), 14 ( G and H

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: Loss of function of PGE2 signaling in MuSCs impairs muscle regeneration and strength. ( A–H ) TAs of Pax7-specific EP4 conditional knockout mice ( Pax7 CreERT2 ;EP4 f/f , EP4 cKO) treated with tamoxifen (TAM) were assayed at 7 ( C and E ), 14 ( G and H

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Knock-Out, Mouse Assay

    EP4 mediates PGE2 signaling in MuSCs. ( A ) Expression of prostaglandin receptors ( Ptger 1–4 ) by MuSCs after 24-h treatment with vehicle or PGE2 ( n = 3 mice with two technical replicates). ( B ) cAMP levels in MuSCs after 1-h PGE2 treatment ( n = 6

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Prostaglandin E2 is essential for efficacious skeletal muscle stem-cell function, augmenting regeneration and strength

    doi: 10.1073/pnas.1705420114

    Figure Lengend Snippet: EP4 mediates PGE2 signaling in MuSCs. ( A ) Expression of prostaglandin receptors ( Ptger 1–4 ) by MuSCs after 24-h treatment with vehicle or PGE2 ( n = 3 mice with two technical replicates). ( B ) cAMP levels in MuSCs after 1-h PGE2 treatment ( n = 6

    Article Snippet: For PGE2, EP4 receptor antagonist treatment studies, we added 1–200 ng/mL Prostaglandin E2 (Cayman Chemical) (unless specified in the figure legends, 10 ng/mL was the standard concentration used), and/or 1 µM EP4 antagonist (ONO-AE3-208, Cayman Chemical) to the MuSCs cultured on collagen-coated dishes for the first 24 h. The cells were then trypsinized and cells reseeded onto hydrogels for an additional 6 d of culture.

    Techniques: Expressing, Mouse Assay

    SUCNR1 Expression Is Necessary for the Anti-inflammatory Effect of NSCs on Type 1 Pro-inflammatory Mφ In Vitro (A) Heatmap showing the microarray expression profile of the 50 most upregulated genes in NSCs after treatment with succinate. Data are shown as Z scores. (B) qRT-PCR independent validation of Ptgs2 expression as in (A). Data are calculated relative to Actb and shown as mean fold change (±SEM) versus untreated cells over n ≥ 3 independent experiments per condition. (C) PGE2 secretion following 1 hr treatment with succinate ± pre-treatment with the selective PTGS2 blocker SC-58125. Data are mean values (±SEM) over n ≥ 3 independent experiments per condition. (D) PGE2 secretion by hiNSCs treated with succinate ± pre-treatment with either SC-58125 or 4c . Data are mean values (±SEM) over n ≥ 3 independent experiments per condition. (E) PGE2 secretion in Mφ co-cultures. Data are mean values (±SEM) over n ≥ 3 independent experiments per condition. (F) Il1b expression relative to Actb in Mφ co-cultures. Data are mean fold change versus Mφ LPS (±SEM) from n ≥ 3 independent experiments per condition. (G) XF assay of the OCR of Mφ as in (E) and (F). Data are normalized on total protein content and expressed as mean values (±SEM) over n ≥ 3 independent experiments per condition. (H) Il1b expression relative to Actb of Mφ co-cultures with hiNSCs. Data are mean fold change versus Mφ LPS (±SEM) from n ≥ 3 independent experiments per condition. (I) XF assay showing the OCR of Mφ as in (H). Data are normalized on total protein content and expressed as mean values (±SEM) over n ≥ 2 independent experiments per condition. (J) PGE2 secretion in Mφ co-cultures as in (H) and (I). Data are mean values (±SEM) over n ≥ 3 independent experiments per condition. ∗ p ≤ 0.05 versus untreated cells (B); ∗ p ≤ 0.05 and ∗∗ p ≤ 0.01 (C and D); ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001 versus Mφ LPS (E–J); # p ≤ 0.05 and ## p ≤ 0.01 versus Mφ LPS -NSCs (E–G) or versus Mφ LPS -hiNSCs (H and I). See also Figure S4 and Table S4 .

    Journal: Cell Stem Cell

    Article Title: Macrophage-Derived Extracellular Succinate Licenses Neural Stem Cells to Suppress Chronic Neuroinflammation

    doi: 10.1016/j.stem.2018.01.020

    Figure Lengend Snippet: SUCNR1 Expression Is Necessary for the Anti-inflammatory Effect of NSCs on Type 1 Pro-inflammatory Mφ In Vitro (A) Heatmap showing the microarray expression profile of the 50 most upregulated genes in NSCs after treatment with succinate. Data are shown as Z scores. (B) qRT-PCR independent validation of Ptgs2 expression as in (A). Data are calculated relative to Actb and shown as mean fold change (±SEM) versus untreated cells over n ≥ 3 independent experiments per condition. (C) PGE2 secretion following 1 hr treatment with succinate ± pre-treatment with the selective PTGS2 blocker SC-58125. Data are mean values (±SEM) over n ≥ 3 independent experiments per condition. (D) PGE2 secretion by hiNSCs treated with succinate ± pre-treatment with either SC-58125 or 4c . Data are mean values (±SEM) over n ≥ 3 independent experiments per condition. (E) PGE2 secretion in Mφ co-cultures. Data are mean values (±SEM) over n ≥ 3 independent experiments per condition. (F) Il1b expression relative to Actb in Mφ co-cultures. Data are mean fold change versus Mφ LPS (±SEM) from n ≥ 3 independent experiments per condition. (G) XF assay of the OCR of Mφ as in (E) and (F). Data are normalized on total protein content and expressed as mean values (±SEM) over n ≥ 3 independent experiments per condition. (H) Il1b expression relative to Actb of Mφ co-cultures with hiNSCs. Data are mean fold change versus Mφ LPS (±SEM) from n ≥ 3 independent experiments per condition. (I) XF assay showing the OCR of Mφ as in (H). Data are normalized on total protein content and expressed as mean values (±SEM) over n ≥ 2 independent experiments per condition. (J) PGE2 secretion in Mφ co-cultures as in (H) and (I). Data are mean values (±SEM) over n ≥ 3 independent experiments per condition. ∗ p ≤ 0.05 versus untreated cells (B); ∗ p ≤ 0.05 and ∗∗ p ≤ 0.01 (C and D); ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001 versus Mφ LPS (E–J); # p ≤ 0.05 and ## p ≤ 0.01 versus Mφ LPS -NSCs (E–G) or versus Mφ LPS -hiNSCs (H and I). See also Figure S4 and Table S4 .

    Article Snippet: All PGE2 levels were determined using PGE2 ELISA kit (Caymanchem) following the manufacturer’s instructions.

    Techniques: Expressing, In Vitro, Microarray, Quantitative RT-PCR, XF Assay

    Transplantation of Sucnr1 Loss-of-Function NSCs Shows Impaired Ability to Ameliorate Chronic Neuroinflammation In Vivo (A) Behavioral outcome of EAE mice. Data are mean EAE score (±SEM) from n ≥ 5 mice/group. (B and C) Flow-cytometry-based ex vivo quantification of the expression levels of type 1 inflammatory (CD80) and anti-inflammatory (MRC1) markers in CX3CR1 + microglial cells (B) and CCR2 + monocyte-derived infiltrating macrophages (C) at 30 dpt. Quantitative data are shown on the left, whereas representative density plots are shown on the right. Data are min to max % of marker-positive cells from n ≥ 4 pools of mice/group. (D and E) Pathological outcomes of experiments as in (A). Data are mean % Bielschowsky negative-stained axonal loss (D) or LFB negative-stained demyelinated (E) areas/spinal cord section (±SEM) from n ≥ 4 mice/group. The scale bars represent 400 μm. (F) PGE2 levels in the CSF and plasma of EAE mice at 30 dpt. Data are mean values (±SEM) from n ≥ 3 samples/group. (G) Succinate levels in the CSF and plasma of EAE mice at 30 dpt. Data are mean values (±SEM) from n ≥ 4 mice/group. Kruskal-Wallis followed by Mann-Whitney post-test is shown. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001 versus PBS; # p ≤ 0.05 versus NSCs. See also Figure S6 .

    Journal: Cell Stem Cell

    Article Title: Macrophage-Derived Extracellular Succinate Licenses Neural Stem Cells to Suppress Chronic Neuroinflammation

    doi: 10.1016/j.stem.2018.01.020

    Figure Lengend Snippet: Transplantation of Sucnr1 Loss-of-Function NSCs Shows Impaired Ability to Ameliorate Chronic Neuroinflammation In Vivo (A) Behavioral outcome of EAE mice. Data are mean EAE score (±SEM) from n ≥ 5 mice/group. (B and C) Flow-cytometry-based ex vivo quantification of the expression levels of type 1 inflammatory (CD80) and anti-inflammatory (MRC1) markers in CX3CR1 + microglial cells (B) and CCR2 + monocyte-derived infiltrating macrophages (C) at 30 dpt. Quantitative data are shown on the left, whereas representative density plots are shown on the right. Data are min to max % of marker-positive cells from n ≥ 4 pools of mice/group. (D and E) Pathological outcomes of experiments as in (A). Data are mean % Bielschowsky negative-stained axonal loss (D) or LFB negative-stained demyelinated (E) areas/spinal cord section (±SEM) from n ≥ 4 mice/group. The scale bars represent 400 μm. (F) PGE2 levels in the CSF and plasma of EAE mice at 30 dpt. Data are mean values (±SEM) from n ≥ 3 samples/group. (G) Succinate levels in the CSF and plasma of EAE mice at 30 dpt. Data are mean values (±SEM) from n ≥ 4 mice/group. Kruskal-Wallis followed by Mann-Whitney post-test is shown. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01, and ∗∗∗ p ≤ 0.001 versus PBS; # p ≤ 0.05 versus NSCs. See also Figure S6 .

    Article Snippet: All PGE2 levels were determined using PGE2 ELISA kit (Caymanchem) following the manufacturer’s instructions.

    Techniques: Transplantation Assay, In Vivo, Mouse Assay, Flow Cytometry, Cytometry, Ex Vivo, Expressing, Derivative Assay, Marker, Staining, MANN-WHITNEY

    Effect of banana flower extract on inflammation in BPH-1 cells. A: BPH-1 cells were treated with vehicle, or banana flower extract for24 h at the indicated concentration. Culture supernatants were analyzed for secreted prostaglandin E2 (PGE2). The data are the mean±SEM ofthree independent experiments. Significantly different at *p

    Journal: In Vivo

    Article Title: Banana Flower Extract Suppresses Benign Prostatic Hyperplasia by Regulating the Inflammatory Response and Inducing G1 Cell-cycle Arrest

    doi: 10.21873/invivo.11389

    Figure Lengend Snippet: Effect of banana flower extract on inflammation in BPH-1 cells. A: BPH-1 cells were treated with vehicle, or banana flower extract for24 h at the indicated concentration. Culture supernatants were analyzed for secreted prostaglandin E2 (PGE2). The data are the mean±SEM ofthree independent experiments. Significantly different at *p

    Article Snippet: Overnight, the medium was refreshed and a series of concentrations of banana flower extract were added to the medium for 24 h. According to the manufacturer’s instructions, culture supernatants were analyzed using PGE2 Enzyme Immunoassay Kit (Cayman Chem., Ann Arbor, MI, USA).

    Techniques: Concentration Assay

    Effect of Equol on LPS-induced NF-κB activation and pro-inflammatory cytokines (TNF-α, IL-6) and PGE-2 secretion in BV-2 cells. BV-2 microglial cells were pretreated with 1, 5, 10, or 20 μM of Equol for 30 min and stimulated with LPS (100 ng/mL) for 1 h for NF-κB related proteins and 24 h for secreted cytokines measurement. Nuclear extracts were prepared using a nuclear extraction kit. Expression of NF-κB, IκB, and p-IκB were measured by western blot. ( A , B ) Protein levels of IκB and p-IκB and their band intensity, respectively; ( C , D ) NF-κB expression and its densitometric analysis. α-Tubulin was used as the loading control. The culture media was subsequently collected to measure the quantity of PGE-2, TNF-α, and IL-6 released by the cells; ( E – G ) secretion of TNF-α, IL-6 and PGE-2 in supernatant was measured using ELISA assay kit. Data are presented as mean ± SEM of three independent experiments performed in triplicate. ** p

    Journal: Nutrients

    Article Title: Equol, a Dietary Daidzein Gut Metabolite Attenuates Microglial Activation and Potentiates Neuroprotection In Vitro

    doi: 10.3390/nu9030207

    Figure Lengend Snippet: Effect of Equol on LPS-induced NF-κB activation and pro-inflammatory cytokines (TNF-α, IL-6) and PGE-2 secretion in BV-2 cells. BV-2 microglial cells were pretreated with 1, 5, 10, or 20 μM of Equol for 30 min and stimulated with LPS (100 ng/mL) for 1 h for NF-κB related proteins and 24 h for secreted cytokines measurement. Nuclear extracts were prepared using a nuclear extraction kit. Expression of NF-κB, IκB, and p-IκB were measured by western blot. ( A , B ) Protein levels of IκB and p-IκB and their band intensity, respectively; ( C , D ) NF-κB expression and its densitometric analysis. α-Tubulin was used as the loading control. The culture media was subsequently collected to measure the quantity of PGE-2, TNF-α, and IL-6 released by the cells; ( E – G ) secretion of TNF-α, IL-6 and PGE-2 in supernatant was measured using ELISA assay kit. Data are presented as mean ± SEM of three independent experiments performed in triplicate. ** p

    Article Snippet: PGE-2 was measured using a competitive ELISA (Cayman Chemical, Ann Arbor, MI, USA) as described previously [ ].

    Techniques: Activation Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

    Inhibition of PGE2 production in AGS cells after exposure to linoleic acid. After 96 h incubation with linoleic acid, the PGE2 accumulation in the medium was determined by an ELISA kit. Data are expressed as mean±SD of three independent experiments. Significance was determined by Student’s t-test ( * P

    Journal: Journal of Cancer Prevention

    Article Title: Linoleic Acid-Induced Growth Inhibition of Human Gastric Epithelial Adenocarcinoma AGS Cells is Associated with Down-Regulation of Prostaglandin E2 Synthesis and Telomerase Activity

    doi:

    Figure Lengend Snippet: Inhibition of PGE2 production in AGS cells after exposure to linoleic acid. After 96 h incubation with linoleic acid, the PGE2 accumulation in the medium was determined by an ELISA kit. Data are expressed as mean±SD of three independent experiments. Significance was determined by Student’s t-test ( * P

    Article Snippet: Measurement of PGE2 production To measure the quantity of PGE2 generated by AGS cells, medium from the cultures under the same conditions was collected and the quantity of PGE2 production was measured using a PGE2 enzyme-linked immunosorbent assay (ELISA) kit (Cayman Chemical Co., Ann Arbor, MI, USA).

    Techniques: Inhibition, Incubation, Enzyme-linked Immunosorbent Assay

    NK-92MI cell migration and subsequent cytotoxicity toward A549/GFP spheroids. NK-92MI (NK) cell migration (blue) was evaluated in the presence and absence of stromal-cell derived factor-1 (SDF-1α) and/or prostaglandin E2 (PGE) in the medium. Percent migration (left axis) was calculated as a ratio of cells in basolateral chamber compared with total cells seeded in insert. Cytotoxicity of A549/GFP cells (green) was evaluated after exposure to NK-92MI (NK) cells in the presence and absence of SDF-1α and/or PGE in the medium. Percent viability (right axis) was calculated via flow cytometry by enumerating GFP-positive A549 cells relative to spheroids without effector cells. Data are shown as the average of two independent studies, n = 24 wells. * p

    Journal: Frontiers in Immunology

    Article Title: A Novel Three-Dimensional Immune Oncology Model for High-Throughput Testing of Tumoricidal Activity

    doi: 10.3389/fimmu.2018.00857

    Figure Lengend Snippet: NK-92MI cell migration and subsequent cytotoxicity toward A549/GFP spheroids. NK-92MI (NK) cell migration (blue) was evaluated in the presence and absence of stromal-cell derived factor-1 (SDF-1α) and/or prostaglandin E2 (PGE) in the medium. Percent migration (left axis) was calculated as a ratio of cells in basolateral chamber compared with total cells seeded in insert. Cytotoxicity of A549/GFP cells (green) was evaluated after exposure to NK-92MI (NK) cells in the presence and absence of SDF-1α and/or PGE in the medium. Percent viability (right axis) was calculated via flow cytometry by enumerating GFP-positive A549 cells relative to spheroids without effector cells. Data are shown as the average of two independent studies, n = 24 wells. * p

    Article Snippet: NK92-MI cells were stained as previously described while simultaneously being treated with 5.7 µM prostaglandin E2 (PGE2 ; Tocris Cat. No. 2296) dissolved in IMDM or vehicle control (IMDM) for 1 h. HTS Transwell-96-Well Permeable Supports were placed in 96-well spheroid plates (a schematic is shown in Figure ).

    Techniques: Migration, Derivative Assay, Flow Cytometry, Cytometry

    Multivariate data analyses of factors responsible for vascular dysfunction. (A) Scores scatter plot t1 vs. t2 resulting after applying PCA to endometrial expression of various angiogenic and vasoactive factors of controls (green) and IRSM (blue), (B) scores scatter plot t1 vs t2 resulting after applying PLS-DA to endometrial expression of various angiogenic and vasoactive factors of controls (green) and IRSM (blue), (C) Results from the permutation test for the IRSM group suggest a valid PLS-DA model. The vertical axis is the R2 and Q2 values of each model and the horizontal axis shows the correlation between the permuted class vectors and the original class vector. The original class has the correlation 1.0 with itself, defining the high point on the horizontal axis. All R2 and Q2 values calculated from the permuted data are lower than the original model in the validation plot. Y-axis intercepts: R2= (0.0, 0.0189), Q2= (0.0, −0.123). (D) Loading scatter plot indicates factors IL-1β, TNF-α, IFN-γ, TGF-β1, PGE2 are upregulated in IRSM and factors IL-2, IL-4, IL-6, IL-8, IL-10, VEGF, ADM, eNOS, NO are upregulated in controls.

    Journal: PLoS ONE

    Article Title: Identification of Key Contributory Factors Responsible for Vascular Dysfunction in Idiopathic Recurrent Spontaneous Miscarriage

    doi: 10.1371/journal.pone.0080940

    Figure Lengend Snippet: Multivariate data analyses of factors responsible for vascular dysfunction. (A) Scores scatter plot t1 vs. t2 resulting after applying PCA to endometrial expression of various angiogenic and vasoactive factors of controls (green) and IRSM (blue), (B) scores scatter plot t1 vs t2 resulting after applying PLS-DA to endometrial expression of various angiogenic and vasoactive factors of controls (green) and IRSM (blue), (C) Results from the permutation test for the IRSM group suggest a valid PLS-DA model. The vertical axis is the R2 and Q2 values of each model and the horizontal axis shows the correlation between the permuted class vectors and the original class vector. The original class has the correlation 1.0 with itself, defining the high point on the horizontal axis. All R2 and Q2 values calculated from the permuted data are lower than the original model in the validation plot. Y-axis intercepts: R2= (0.0, 0.0189), Q2= (0.0, −0.123). (D) Loading scatter plot indicates factors IL-1β, TNF-α, IFN-γ, TGF-β1, PGE2 are upregulated in IRSM and factors IL-2, IL-4, IL-6, IL-8, IL-10, VEGF, ADM, eNOS, NO are upregulated in controls.

    Article Snippet: ELISA Tissue homogenates containing protein concentration of 30 μg/ml were used for estimating the level of IL-1β, TNF-α, IFN-γ, TGF-β1, IL-10, -2, -6, -8, VEGF, eNOS and ADM. VEGF and PGE2 were assessed using anti-human rabbit polyclonal antibody to VEGF (ab9570) and PGE2 (ab-2318; Abcam, Cambridge, UK), respectively employing quantitative direct enzyme immunoassay technique.

    Techniques: Expressing, Plasmid Preparation

    Ectopic Expression of miR-411 Induced Cell Cycles Arrest in G2/M Phase by Up-regulating p21 Protein in Human BC Cells (A) Cell lysates from the indicated cells were subjected to western blot to analyze expression of p21, cyclin B1, cyclin A2, and cyclin E2. β-actin was used as an internal control. (B) Western blot was used to determine the expression of p21 in T24 (miR-411/HA-MLLT11) and UMUC3 (miR-411/HA-MLLT11) cells in comparison to their vector control transfectants. β-actin was used as an internal control. (C) UMUC3 (miR-411) cells stably transfected with shRNA targeting human p21 or its nonsense control plasmid. The cell lysates from the indicated transfectants were subjected to western blot to assess the knockdown efficiency of the p21 protein. (D) UMUC3 (miR-411/nonsense), UMUC3 (miR-411/shp21 No. 3), and UMUC3 (miR-411/shp21 No. 6) cells were subjected to soft-agar assay to determine the effect of p21 on miR-411-induced inhibition of anchorage-independent growth. The images of colonies from the indicated cells were captured under microscopy after 3 weeks of incubation. (E) The number of colonies was scored and presented as colonies per 10,000 cells. The symbol (*) indicates a significant increase in comparison with UMUC3 (miR-411/Nonsense) transfectants (*p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: MicroRNA-411 Downregulation Enhances Tumor Growth by Upregulating MLLT11 Expression in Human Bladder Cancer

    doi: 10.1016/j.omtn.2018.03.003

    Figure Lengend Snippet: Ectopic Expression of miR-411 Induced Cell Cycles Arrest in G2/M Phase by Up-regulating p21 Protein in Human BC Cells (A) Cell lysates from the indicated cells were subjected to western blot to analyze expression of p21, cyclin B1, cyclin A2, and cyclin E2. β-actin was used as an internal control. (B) Western blot was used to determine the expression of p21 in T24 (miR-411/HA-MLLT11) and UMUC3 (miR-411/HA-MLLT11) cells in comparison to their vector control transfectants. β-actin was used as an internal control. (C) UMUC3 (miR-411) cells stably transfected with shRNA targeting human p21 or its nonsense control plasmid. The cell lysates from the indicated transfectants were subjected to western blot to assess the knockdown efficiency of the p21 protein. (D) UMUC3 (miR-411/nonsense), UMUC3 (miR-411/shp21 No. 3), and UMUC3 (miR-411/shp21 No. 6) cells were subjected to soft-agar assay to determine the effect of p21 on miR-411-induced inhibition of anchorage-independent growth. The images of colonies from the indicated cells were captured under microscopy after 3 weeks of incubation. (E) The number of colonies was scored and presented as colonies per 10,000 cells. The symbol (*) indicates a significant increase in comparison with UMUC3 (miR-411/Nonsense) transfectants (*p

    Article Snippet: The antibodies against cyclin A2 (No. 4656), cyclin B1 (No. 12231), cyclin E2 (No. 4132), EGFR (No. 4267S), and ATG3 (No. 3415) were obtained from Cell Signaling Technology (Boston, MA, USA).

    Techniques: Expressing, Western Blot, Plasmid Preparation, Stable Transfection, Transfection, shRNA, Soft Agar Assay, Inhibition, Microscopy, Incubation

    Germline-encoded clone strongly reacted to xenobiotics A) AA sequences of cross-reactive clones S1-8 and S1-9 aligned with IGHV 3-30 germline sequence. B) Schematic representation of conversion of germline IGHV3-30-reverted antibody generation. C) A side-by-side comparison of antigen reactivities against PDC-E2 and 2OA-BSA using immunoblots. Equal concentrations of mAbs were used to probe PDC-E2 and 2OA-BSA.

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Autoreactive Monoclonal Antibodies from Patients with Primary Biliary Cholangitis Recognize Environmental Xenobiotics

    doi: 10.1002/hep.29245

    Figure Lengend Snippet: Germline-encoded clone strongly reacted to xenobiotics A) AA sequences of cross-reactive clones S1-8 and S1-9 aligned with IGHV 3-30 germline sequence. B) Schematic representation of conversion of germline IGHV3-30-reverted antibody generation. C) A side-by-side comparison of antigen reactivities against PDC-E2 and 2OA-BSA using immunoblots. Equal concentrations of mAbs were used to probe PDC-E2 and 2OA-BSA.

    Article Snippet: Among these 104 single mAbs, 32 (30.8%) reacted to PDC-E2, including 20 reacted to PDC-E2 only and 12 cross-reacted with PDC-E2 and xenobiotics ( ).

    Techniques: Clone Assay, Sequencing, Western Blot

    Antigen specificity and cross-reactivity of mAbs against PDC-E2, 2OA-BSA and SAc-BSA. A) Immunoblot assay was used to identify tantigen specificity and cross-reactivity. mAbs cross-reacting with PDC-E2, 2OA-BSA and SAc-BSA were detected in all three subjects (S1, S2 and S3). Culture media only were served as negative controls (N ctrl); PBC patient sera diluted from 1:1,000 to 1:200,000 served as positive controls (P ctrl). B) The frequencies of PDC-E2 negative, PDC-E2 reactive and PDC-E2/xenobiotics cross-reactive clones. Antibody isotype distribution were examined from 104 randomly selected clones and screened for antigen specificity and cross-reactivity against PDC-E2, 2OA-BSA and SAc-BSA (IgA, n=63; IgM, n=22; IgG, n=19). The numbers in pie charts reflect the number of PDC-E2 negative, PDC-E2 reactive and PDC-E2/xenobiotics cross-reactive clones, respectively.

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Autoreactive Monoclonal Antibodies from Patients with Primary Biliary Cholangitis Recognize Environmental Xenobiotics

    doi: 10.1002/hep.29245

    Figure Lengend Snippet: Antigen specificity and cross-reactivity of mAbs against PDC-E2, 2OA-BSA and SAc-BSA. A) Immunoblot assay was used to identify tantigen specificity and cross-reactivity. mAbs cross-reacting with PDC-E2, 2OA-BSA and SAc-BSA were detected in all three subjects (S1, S2 and S3). Culture media only were served as negative controls (N ctrl); PBC patient sera diluted from 1:1,000 to 1:200,000 served as positive controls (P ctrl). B) The frequencies of PDC-E2 negative, PDC-E2 reactive and PDC-E2/xenobiotics cross-reactive clones. Antibody isotype distribution were examined from 104 randomly selected clones and screened for antigen specificity and cross-reactivity against PDC-E2, 2OA-BSA and SAc-BSA (IgA, n=63; IgM, n=22; IgG, n=19). The numbers in pie charts reflect the number of PDC-E2 negative, PDC-E2 reactive and PDC-E2/xenobiotics cross-reactive clones, respectively.

    Article Snippet: Among these 104 single mAbs, 32 (30.8%) reacted to PDC-E2, including 20 reacted to PDC-E2 only and 12 cross-reacted with PDC-E2 and xenobiotics ( ).

    Techniques: Clone Assay

    Comparison of number of mutations in IGHV regions and IGLV regions in PDC-E2/xenobiotic cross-reactive, PDC-E2 reactive and PDC-E2 negative mAbs A) Comparison of the number of mutations in IGHV, framework (FR) and CDR region of IGHV sequences (upper panels). Comparison of the number of replacement (R) and silent (S) mutations in IGHV and R-mutation in CDR region of IGHV-sequences (lower panels). B) Comparison of the number of mutations in IGLV, FR and CDR region of IGLV sequences (upper panels). Comparison of the number of R-and S-mutation in IGLV, and R-mutation in the CDR region of IGLV-sequences (lower panels).

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Autoreactive Monoclonal Antibodies from Patients with Primary Biliary Cholangitis Recognize Environmental Xenobiotics

    doi: 10.1002/hep.29245

    Figure Lengend Snippet: Comparison of number of mutations in IGHV regions and IGLV regions in PDC-E2/xenobiotic cross-reactive, PDC-E2 reactive and PDC-E2 negative mAbs A) Comparison of the number of mutations in IGHV, framework (FR) and CDR region of IGHV sequences (upper panels). Comparison of the number of replacement (R) and silent (S) mutations in IGHV and R-mutation in CDR region of IGHV-sequences (lower panels). B) Comparison of the number of mutations in IGLV, FR and CDR region of IGLV sequences (upper panels). Comparison of the number of R-and S-mutation in IGLV, and R-mutation in the CDR region of IGLV-sequences (lower panels).

    Article Snippet: Among these 104 single mAbs, 32 (30.8%) reacted to PDC-E2, including 20 reacted to PDC-E2 only and 12 cross-reacted with PDC-E2 and xenobiotics ( ).

    Techniques: Mutagenesis

    CDR1 amino acid (AA) sequence in IGHV 4-61:IGLV 2-14 family is critical for PDC-E2 recognition A) Evolutionary AA replacements were analyzed in six evolutionarily closely related clones, which utilized IGHV4-61 and IGLV2-14. B) 3-D structures of PDC-E2 reactive (S1-3, S1-10, S1-11, S1-12 and S1-13) and a PDC-E2 negative clone (S1-16). Orange arrows indicate the sites of CDRs mutations. Note that 2AA replacements (G→S and I→V) in CDR1 did not affect the reactions to PDC-E2, whereas AA replacement (G→D) located on the surface in CDR1 region resulted in loss of PDC-E2 recognition. C) Illustration of artificial recombinant mAb construction. The IGHV segment was cloned from PDC-E2 positive antibody S1-13, and the IGLV segment was cloned from PDC-E2 negative S1-16. D) The AA replacements in IGHV were more critical for antigen reactivity than that in IGLV. Immunoblotting results demonstrated that PDC-E2 reactivity was comparable between the original clone S1-13 and designer clone “S1-13 IGHV: S1-16 IGLV” (S1-13 IGHV was constructed by pairing IGLV from the PDC-E2 negative clone S1-16).

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Autoreactive Monoclonal Antibodies from Patients with Primary Biliary Cholangitis Recognize Environmental Xenobiotics

    doi: 10.1002/hep.29245

    Figure Lengend Snippet: CDR1 amino acid (AA) sequence in IGHV 4-61:IGLV 2-14 family is critical for PDC-E2 recognition A) Evolutionary AA replacements were analyzed in six evolutionarily closely related clones, which utilized IGHV4-61 and IGLV2-14. B) 3-D structures of PDC-E2 reactive (S1-3, S1-10, S1-11, S1-12 and S1-13) and a PDC-E2 negative clone (S1-16). Orange arrows indicate the sites of CDRs mutations. Note that 2AA replacements (G→S and I→V) in CDR1 did not affect the reactions to PDC-E2, whereas AA replacement (G→D) located on the surface in CDR1 region resulted in loss of PDC-E2 recognition. C) Illustration of artificial recombinant mAb construction. The IGHV segment was cloned from PDC-E2 positive antibody S1-13, and the IGLV segment was cloned from PDC-E2 negative S1-16. D) The AA replacements in IGHV were more critical for antigen reactivity than that in IGLV. Immunoblotting results demonstrated that PDC-E2 reactivity was comparable between the original clone S1-13 and designer clone “S1-13 IGHV: S1-16 IGLV” (S1-13 IGHV was constructed by pairing IGLV from the PDC-E2 negative clone S1-16).

    Article Snippet: Among these 104 single mAbs, 32 (30.8%) reacted to PDC-E2, including 20 reacted to PDC-E2 only and 12 cross-reacted with PDC-E2 and xenobiotics ( ).

    Techniques: Sequencing, Clone Assay, Recombinant, Construct

    Cross-reactive mAbs recognize LA. Reactivity of PDC-E2/xenobiotic cross-reactive clones to lipoic acid (LA). Immunoblotting results demonstrate that PDC-E2/xenobiotic cross-reactive mAbs reacting to LA-RSA, but not RSA. N ctrl, negative control; P ctrl, positive control.

    Journal: Hepatology (Baltimore, Md.)

    Article Title: Autoreactive Monoclonal Antibodies from Patients with Primary Biliary Cholangitis Recognize Environmental Xenobiotics

    doi: 10.1002/hep.29245

    Figure Lengend Snippet: Cross-reactive mAbs recognize LA. Reactivity of PDC-E2/xenobiotic cross-reactive clones to lipoic acid (LA). Immunoblotting results demonstrate that PDC-E2/xenobiotic cross-reactive mAbs reacting to LA-RSA, but not RSA. N ctrl, negative control; P ctrl, positive control.

    Article Snippet: Among these 104 single mAbs, 32 (30.8%) reacted to PDC-E2, including 20 reacted to PDC-E2 only and 12 cross-reacted with PDC-E2 and xenobiotics ( ).

    Techniques: Clone Assay, Negative Control, Positive Control