anti cyclin d1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti cyclin d1
    Primer sequences for quantitative PCR.
    Anti Cyclin D1, 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
    https://www.bioz.com/result/anti cyclin d1/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cyclin d1 - by Bioz Stars, 2024-06
    86/100 stars

    Images

    1) Product Images from "Calcitonin gene‑related peptide alleviates hyperoxia‑induced human alveolar cell injury via the CGRPR/TRPV1/Ca2 + axis"

    Article Title: Calcitonin gene‑related peptide alleviates hyperoxia‑induced human alveolar cell injury via the CGRPR/TRPV1/Ca2 + axis

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2024.13234

    Primer sequences for quantitative PCR.
    Figure Legend Snippet: Primer sequences for quantitative PCR.

    Techniques Used: Sequencing

    The CGRPR/TRPV1 axis regulates the mRNA and protein expression levels of proliferation and apoptosis factors in A549 cells under hyperoxic conditions. (A and B) quantitative PCR revealed the effects of 10 nM CGRP, 100 nM CGRP 8-37 and 10 µM SB-705498 on the expression of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. (C) Western blotting revealed the effects of CGRP and CGRP 8-37 on the expression levels of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. (D) Western blotting revealed the effects of CGRP and SB-705498 on the expression levels of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. Data are presented as the mean ± SD of at least three repeats. *P<0.05, **P<0.01 and ***P<0.001. CGRP, calcitonin gene-related peptide; CGRPR, CGRP receptor; TRPV1, transient receptor potential vanilloid 1; PCNA, proliferating cell nuclear antigen.
    Figure Legend Snippet: The CGRPR/TRPV1 axis regulates the mRNA and protein expression levels of proliferation and apoptosis factors in A549 cells under hyperoxic conditions. (A and B) quantitative PCR revealed the effects of 10 nM CGRP, 100 nM CGRP 8-37 and 10 µM SB-705498 on the expression of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. (C) Western blotting revealed the effects of CGRP and CGRP 8-37 on the expression levels of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. (D) Western blotting revealed the effects of CGRP and SB-705498 on the expression levels of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. Data are presented as the mean ± SD of at least three repeats. *P<0.05, **P<0.01 and ***P<0.001. CGRP, calcitonin gene-related peptide; CGRPR, CGRP receptor; TRPV1, transient receptor potential vanilloid 1; PCNA, proliferating cell nuclear antigen.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Western Blot

    Knockdown of TRPV1 confirms the role of TRPV1 in CGRP-mediated cell proliferation and apoptosis of A549 cells cultured under hyperoxic conditions. (A and B) The mRNA and protein expression levels of TRPV1. For knockdown of TRPV1, three lentiviruses with different interference sequences were used. ShTRPV1-2 was used for subsequent experiments. (C) shTRPV1 attenuated the CGRP-induced increase in proliferation of A549 cells under hyperoxic conditions. (D) shTRPV1 attenuated the effect of CGRP-induced decrease in apoptosis of A549 cells under hyperoxic conditions. (E) Effects of CGRP on Cyclin D1, PCNA, Bcl-2 and Bax expression in A549 shTRPV1 cells cultured under hyperoxic conditions. Data are presented as the mean ± SD of at least three repeats. **P<0.01 and ***P<0.001. TRPV1, transient receptor potential vanilloid 1; CGRP, calcitonin gene-related peptide; m, messenger; sh, short hairpin; PCNA, proliferating cell nuclear antigen; NC, negative controls; ns, not significant; PI, propidium iodine.
    Figure Legend Snippet: Knockdown of TRPV1 confirms the role of TRPV1 in CGRP-mediated cell proliferation and apoptosis of A549 cells cultured under hyperoxic conditions. (A and B) The mRNA and protein expression levels of TRPV1. For knockdown of TRPV1, three lentiviruses with different interference sequences were used. ShTRPV1-2 was used for subsequent experiments. (C) shTRPV1 attenuated the CGRP-induced increase in proliferation of A549 cells under hyperoxic conditions. (D) shTRPV1 attenuated the effect of CGRP-induced decrease in apoptosis of A549 cells under hyperoxic conditions. (E) Effects of CGRP on Cyclin D1, PCNA, Bcl-2 and Bax expression in A549 shTRPV1 cells cultured under hyperoxic conditions. Data are presented as the mean ± SD of at least three repeats. **P<0.01 and ***P<0.001. TRPV1, transient receptor potential vanilloid 1; CGRP, calcitonin gene-related peptide; m, messenger; sh, short hairpin; PCNA, proliferating cell nuclear antigen; NC, negative controls; ns, not significant; PI, propidium iodine.

    Techniques Used: Cell Culture, Expressing

    CGRP/CGRPR regulates TRPV1 via the PLC/PKC pathway in A549 cells cultured under hyperoxic conditions. (A and B) Cell Counting Kit-8 assay revealed that 10 nM CGRP promoted A549 cell proliferation under hyperoxic conditions, and this was inhibited by both 1 µM U-73122 and 200 nM Go6976. (C and D) Western blotting revealed CGRP increased the expression of Cyclin D1, PCNA and Bcl-2 and attenuated the expression of Bax in hyperoxia, whereas both U-73122 and Go6976 reversed the effects of CGRP. (E and F) The densitometric analysis diagram of the western blots. Data are presented as the mean ± SD of at least three repeats. **P<0.01 and ***P<0.001; CGRP, calcitonin gene-related peptide; CGRPR, CGRP receptor; TRPV1, transient receptor potential vanilloid 1; PLC, phospholipase C; PKC, protein kinase C; PCNA, proliferating cell nuclear antigen; ns, not significant.
    Figure Legend Snippet: CGRP/CGRPR regulates TRPV1 via the PLC/PKC pathway in A549 cells cultured under hyperoxic conditions. (A and B) Cell Counting Kit-8 assay revealed that 10 nM CGRP promoted A549 cell proliferation under hyperoxic conditions, and this was inhibited by both 1 µM U-73122 and 200 nM Go6976. (C and D) Western blotting revealed CGRP increased the expression of Cyclin D1, PCNA and Bcl-2 and attenuated the expression of Bax in hyperoxia, whereas both U-73122 and Go6976 reversed the effects of CGRP. (E and F) The densitometric analysis diagram of the western blots. Data are presented as the mean ± SD of at least three repeats. **P<0.01 and ***P<0.001; CGRP, calcitonin gene-related peptide; CGRPR, CGRP receptor; TRPV1, transient receptor potential vanilloid 1; PLC, phospholipase C; PKC, protein kinase C; PCNA, proliferating cell nuclear antigen; ns, not significant.

    Techniques Used: Cell Culture, Cell Counting, Western Blot, Expressing

    cyclin d1 cat no 55506s antibodies  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc cyclin d1 cat no 55506s antibodies
    Cyclin D1 Cat No 55506s Antibodies, 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
    https://www.bioz.com/result/cyclin d1 cat no 55506s antibodies/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cyclin d1 cat no 55506s antibodies - by Bioz Stars, 2024-06
    86/100 stars

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    antibodies against cyclin d1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc antibodies against cyclin d1
    A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of <t>cyclin</t> <t>D1</t> protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).
    Antibodies Against Cyclin D1, 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
    https://www.bioz.com/result/antibodies against cyclin d1/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    Images

    1) Product Images from "Syrosingopine and UK5099 synergistically suppress non-small cell lung cancer by activating the integrated stress response"

    Article Title: Syrosingopine and UK5099 synergistically suppress non-small cell lung cancer by activating the integrated stress response

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-024-06821-4

    A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of cyclin D1 protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).
    Figure Legend Snippet: A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of cyclin D1 protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).

    Techniques Used: Cell Cycle Assay, Western Blot, Scratch Wound Assay Assay, Staining

    cyclin d1  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc cyclin d1
    Cyclin D1, 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
    https://www.bioz.com/result/cyclin d1/product/Cell Signaling Technology Inc
    Average 86 stars, based on 1 article reviews
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    cyclin d1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc cyclin d1
    Circ6834 overexpression suppresses NSCLC growth and metastasis in vitro and in vivo. A The efficiency of circ6834 overexpression in A549 cells. B-E, G Cell growth curves ( B ), colony formation assays ( C ), flow cytometric analyses of cell apoptosis ( D ) and cell cycle ( E ), and Transwell migration and Matrigel invasion assays ( G ) for EGFP and circ6834-overexpressing A549 cells. F, H Western blot analyses for the expression of <t>Cyclin</t> <t>D1,</t> Bcl-2 ( F ), and EMT markers ( H ) in EGFP and circ6834-overexpressing A549 cells. I The effects of circ6834 overexpression on the sensitivity of A549 cells to DDP and OXA treatment. J Images of tumors from mice in the EGFP and circ6834-overexpressing groups. K-L H&E, Ki-67 ( K ) and TUNEL ( L ) staining of mouse tumor tissues from the EGFP and circ6834-overexpressing groups. M Representative images and IHC staining of lung tissues with metastatic modules from mice in the EGFP and circ6834-overexpressing groups
    Cyclin D1, 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
    https://www.bioz.com/result/cyclin d1/product/Cell Signaling Technology Inc
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    Images

    1) Product Images from "Circ6834 suppresses non-small cell lung cancer progression by destabilizing ANHAK and regulating miR-873-5p/TXNIP axis"

    Article Title: Circ6834 suppresses non-small cell lung cancer progression by destabilizing ANHAK and regulating miR-873-5p/TXNIP axis

    Journal: Molecular Cancer

    doi: 10.1186/s12943-024-02038-3

    Circ6834 overexpression suppresses NSCLC growth and metastasis in vitro and in vivo. A The efficiency of circ6834 overexpression in A549 cells. B-E, G Cell growth curves ( B ), colony formation assays ( C ), flow cytometric analyses of cell apoptosis ( D ) and cell cycle ( E ), and Transwell migration and Matrigel invasion assays ( G ) for EGFP and circ6834-overexpressing A549 cells. F, H Western blot analyses for the expression of Cyclin D1, Bcl-2 ( F ), and EMT markers ( H ) in EGFP and circ6834-overexpressing A549 cells. I The effects of circ6834 overexpression on the sensitivity of A549 cells to DDP and OXA treatment. J Images of tumors from mice in the EGFP and circ6834-overexpressing groups. K-L H&E, Ki-67 ( K ) and TUNEL ( L ) staining of mouse tumor tissues from the EGFP and circ6834-overexpressing groups. M Representative images and IHC staining of lung tissues with metastatic modules from mice in the EGFP and circ6834-overexpressing groups
    Figure Legend Snippet: Circ6834 overexpression suppresses NSCLC growth and metastasis in vitro and in vivo. A The efficiency of circ6834 overexpression in A549 cells. B-E, G Cell growth curves ( B ), colony formation assays ( C ), flow cytometric analyses of cell apoptosis ( D ) and cell cycle ( E ), and Transwell migration and Matrigel invasion assays ( G ) for EGFP and circ6834-overexpressing A549 cells. F, H Western blot analyses for the expression of Cyclin D1, Bcl-2 ( F ), and EMT markers ( H ) in EGFP and circ6834-overexpressing A549 cells. I The effects of circ6834 overexpression on the sensitivity of A549 cells to DDP and OXA treatment. J Images of tumors from mice in the EGFP and circ6834-overexpressing groups. K-L H&E, Ki-67 ( K ) and TUNEL ( L ) staining of mouse tumor tissues from the EGFP and circ6834-overexpressing groups. M Representative images and IHC staining of lung tissues with metastatic modules from mice in the EGFP and circ6834-overexpressing groups

    Techniques Used: Over Expression, In Vitro, In Vivo, Migration, Western Blot, Expressing, TUNEL Assay, Staining, Immunohistochemistry

    cyclin d1 ta801655  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc cyclin d1 ta801655
    Cyclin D1 Ta801655, 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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    cyclin d1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc cyclin d1
    Cyclin D1, 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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    rabbit anti cyclin d1  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti cyclin d1
    Rabbit Anti Cyclin D1, 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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    cyclin d1  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc cyclin d1
    Cyclin D1, 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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    anti cyclin d1  (Cell Signaling Technology Inc)


    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
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    Cell Signaling Technology Inc anti cyclin d1
    Anti Cyclin D1, 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
    https://www.bioz.com/result/anti cyclin d1/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc anti cyclin d1
    Primer sequences for quantitative PCR.
    Anti Cyclin D1, 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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    Cell Signaling Technology Inc cyclin d1 cat no 55506s antibodies
    Primer sequences for quantitative PCR.
    Cyclin D1 Cat No 55506s Antibodies, 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
    https://www.bioz.com/result/cyclin d1 cat no 55506s antibodies/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc antibodies against cyclin d1
    A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of <t>cyclin</t> <t>D1</t> protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).
    Antibodies Against Cyclin D1, 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
    https://www.bioz.com/result/antibodies against cyclin d1/product/Cell Signaling Technology Inc
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    Cell Signaling Technology Inc cyclin d1
    A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of <t>cyclin</t> <t>D1</t> protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).
    Cyclin D1, 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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    Cell Signaling Technology Inc cyclin d1 ta801655
    A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of <t>cyclin</t> <t>D1</t> protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).
    Cyclin D1 Ta801655, 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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    Cell Signaling Technology Inc rabbit anti cyclin d1
    A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of <t>cyclin</t> <t>D1</t> protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).
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    Primer sequences for quantitative PCR.

    Journal: Molecular Medicine Reports

    Article Title: Calcitonin gene‑related peptide alleviates hyperoxia‑induced human alveolar cell injury via the CGRPR/TRPV1/Ca2 + axis

    doi: 10.3892/mmr.2024.13234

    Figure Lengend Snippet: Primer sequences for quantitative PCR.

    Article Snippet: Blots were blocked for 10 min at room temperature using rapid blot blocking buffer (cat. no. P30500; New Cell & Molecular Biotech Co., Ltd.), and then incubated at 4°C overnight with the following specific primary antibodies: anti-TRPV1 (1:500), anti-CGRPR (1:500), anti-PCNA (1:1,000; cat. no. 13110; Cell Signaling Technology, Inc.), anti-Cyclin D1 (1:1,000; cat. no. 55506; Cell Signaling Technology, Inc.), anti-Bcl-2 (1:1,000; cat. no. 60178-1-Ig, ProteinTech Group, Inc.), anti-Bax (1:5,000; cat. no. 60267-1-Ig; ProteinTech Group, Inc.), anti-β-actin (1:1,000; cat. no. 4970; Cell Signaling Technology, Inc.), or anti-GAPDH (1:1,000; cat. no. 2118; Cell Signaling Technology, Inc.).

    Techniques: Sequencing

    The CGRPR/TRPV1 axis regulates the mRNA and protein expression levels of proliferation and apoptosis factors in A549 cells under hyperoxic conditions. (A and B) quantitative PCR revealed the effects of 10 nM CGRP, 100 nM CGRP 8-37 and 10 µM SB-705498 on the expression of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. (C) Western blotting revealed the effects of CGRP and CGRP 8-37 on the expression levels of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. (D) Western blotting revealed the effects of CGRP and SB-705498 on the expression levels of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. Data are presented as the mean ± SD of at least three repeats. *P<0.05, **P<0.01 and ***P<0.001. CGRP, calcitonin gene-related peptide; CGRPR, CGRP receptor; TRPV1, transient receptor potential vanilloid 1; PCNA, proliferating cell nuclear antigen.

    Journal: Molecular Medicine Reports

    Article Title: Calcitonin gene‑related peptide alleviates hyperoxia‑induced human alveolar cell injury via the CGRPR/TRPV1/Ca2 + axis

    doi: 10.3892/mmr.2024.13234

    Figure Lengend Snippet: The CGRPR/TRPV1 axis regulates the mRNA and protein expression levels of proliferation and apoptosis factors in A549 cells under hyperoxic conditions. (A and B) quantitative PCR revealed the effects of 10 nM CGRP, 100 nM CGRP 8-37 and 10 µM SB-705498 on the expression of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. (C) Western blotting revealed the effects of CGRP and CGRP 8-37 on the expression levels of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. (D) Western blotting revealed the effects of CGRP and SB-705498 on the expression levels of Cyclin D1, PCNA, Bcl-2 and Bax in A549 cells cultured under hyperoxic conditions. Data are presented as the mean ± SD of at least three repeats. *P<0.05, **P<0.01 and ***P<0.001. CGRP, calcitonin gene-related peptide; CGRPR, CGRP receptor; TRPV1, transient receptor potential vanilloid 1; PCNA, proliferating cell nuclear antigen.

    Article Snippet: Blots were blocked for 10 min at room temperature using rapid blot blocking buffer (cat. no. P30500; New Cell & Molecular Biotech Co., Ltd.), and then incubated at 4°C overnight with the following specific primary antibodies: anti-TRPV1 (1:500), anti-CGRPR (1:500), anti-PCNA (1:1,000; cat. no. 13110; Cell Signaling Technology, Inc.), anti-Cyclin D1 (1:1,000; cat. no. 55506; Cell Signaling Technology, Inc.), anti-Bcl-2 (1:1,000; cat. no. 60178-1-Ig, ProteinTech Group, Inc.), anti-Bax (1:5,000; cat. no. 60267-1-Ig; ProteinTech Group, Inc.), anti-β-actin (1:1,000; cat. no. 4970; Cell Signaling Technology, Inc.), or anti-GAPDH (1:1,000; cat. no. 2118; Cell Signaling Technology, Inc.).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Western Blot

    Knockdown of TRPV1 confirms the role of TRPV1 in CGRP-mediated cell proliferation and apoptosis of A549 cells cultured under hyperoxic conditions. (A and B) The mRNA and protein expression levels of TRPV1. For knockdown of TRPV1, three lentiviruses with different interference sequences were used. ShTRPV1-2 was used for subsequent experiments. (C) shTRPV1 attenuated the CGRP-induced increase in proliferation of A549 cells under hyperoxic conditions. (D) shTRPV1 attenuated the effect of CGRP-induced decrease in apoptosis of A549 cells under hyperoxic conditions. (E) Effects of CGRP on Cyclin D1, PCNA, Bcl-2 and Bax expression in A549 shTRPV1 cells cultured under hyperoxic conditions. Data are presented as the mean ± SD of at least three repeats. **P<0.01 and ***P<0.001. TRPV1, transient receptor potential vanilloid 1; CGRP, calcitonin gene-related peptide; m, messenger; sh, short hairpin; PCNA, proliferating cell nuclear antigen; NC, negative controls; ns, not significant; PI, propidium iodine.

    Journal: Molecular Medicine Reports

    Article Title: Calcitonin gene‑related peptide alleviates hyperoxia‑induced human alveolar cell injury via the CGRPR/TRPV1/Ca2 + axis

    doi: 10.3892/mmr.2024.13234

    Figure Lengend Snippet: Knockdown of TRPV1 confirms the role of TRPV1 in CGRP-mediated cell proliferation and apoptosis of A549 cells cultured under hyperoxic conditions. (A and B) The mRNA and protein expression levels of TRPV1. For knockdown of TRPV1, three lentiviruses with different interference sequences were used. ShTRPV1-2 was used for subsequent experiments. (C) shTRPV1 attenuated the CGRP-induced increase in proliferation of A549 cells under hyperoxic conditions. (D) shTRPV1 attenuated the effect of CGRP-induced decrease in apoptosis of A549 cells under hyperoxic conditions. (E) Effects of CGRP on Cyclin D1, PCNA, Bcl-2 and Bax expression in A549 shTRPV1 cells cultured under hyperoxic conditions. Data are presented as the mean ± SD of at least three repeats. **P<0.01 and ***P<0.001. TRPV1, transient receptor potential vanilloid 1; CGRP, calcitonin gene-related peptide; m, messenger; sh, short hairpin; PCNA, proliferating cell nuclear antigen; NC, negative controls; ns, not significant; PI, propidium iodine.

    Article Snippet: Blots were blocked for 10 min at room temperature using rapid blot blocking buffer (cat. no. P30500; New Cell & Molecular Biotech Co., Ltd.), and then incubated at 4°C overnight with the following specific primary antibodies: anti-TRPV1 (1:500), anti-CGRPR (1:500), anti-PCNA (1:1,000; cat. no. 13110; Cell Signaling Technology, Inc.), anti-Cyclin D1 (1:1,000; cat. no. 55506; Cell Signaling Technology, Inc.), anti-Bcl-2 (1:1,000; cat. no. 60178-1-Ig, ProteinTech Group, Inc.), anti-Bax (1:5,000; cat. no. 60267-1-Ig; ProteinTech Group, Inc.), anti-β-actin (1:1,000; cat. no. 4970; Cell Signaling Technology, Inc.), or anti-GAPDH (1:1,000; cat. no. 2118; Cell Signaling Technology, Inc.).

    Techniques: Cell Culture, Expressing

    CGRP/CGRPR regulates TRPV1 via the PLC/PKC pathway in A549 cells cultured under hyperoxic conditions. (A and B) Cell Counting Kit-8 assay revealed that 10 nM CGRP promoted A549 cell proliferation under hyperoxic conditions, and this was inhibited by both 1 µM U-73122 and 200 nM Go6976. (C and D) Western blotting revealed CGRP increased the expression of Cyclin D1, PCNA and Bcl-2 and attenuated the expression of Bax in hyperoxia, whereas both U-73122 and Go6976 reversed the effects of CGRP. (E and F) The densitometric analysis diagram of the western blots. Data are presented as the mean ± SD of at least three repeats. **P<0.01 and ***P<0.001; CGRP, calcitonin gene-related peptide; CGRPR, CGRP receptor; TRPV1, transient receptor potential vanilloid 1; PLC, phospholipase C; PKC, protein kinase C; PCNA, proliferating cell nuclear antigen; ns, not significant.

    Journal: Molecular Medicine Reports

    Article Title: Calcitonin gene‑related peptide alleviates hyperoxia‑induced human alveolar cell injury via the CGRPR/TRPV1/Ca2 + axis

    doi: 10.3892/mmr.2024.13234

    Figure Lengend Snippet: CGRP/CGRPR regulates TRPV1 via the PLC/PKC pathway in A549 cells cultured under hyperoxic conditions. (A and B) Cell Counting Kit-8 assay revealed that 10 nM CGRP promoted A549 cell proliferation under hyperoxic conditions, and this was inhibited by both 1 µM U-73122 and 200 nM Go6976. (C and D) Western blotting revealed CGRP increased the expression of Cyclin D1, PCNA and Bcl-2 and attenuated the expression of Bax in hyperoxia, whereas both U-73122 and Go6976 reversed the effects of CGRP. (E and F) The densitometric analysis diagram of the western blots. Data are presented as the mean ± SD of at least three repeats. **P<0.01 and ***P<0.001; CGRP, calcitonin gene-related peptide; CGRPR, CGRP receptor; TRPV1, transient receptor potential vanilloid 1; PLC, phospholipase C; PKC, protein kinase C; PCNA, proliferating cell nuclear antigen; ns, not significant.

    Article Snippet: Blots were blocked for 10 min at room temperature using rapid blot blocking buffer (cat. no. P30500; New Cell & Molecular Biotech Co., Ltd.), and then incubated at 4°C overnight with the following specific primary antibodies: anti-TRPV1 (1:500), anti-CGRPR (1:500), anti-PCNA (1:1,000; cat. no. 13110; Cell Signaling Technology, Inc.), anti-Cyclin D1 (1:1,000; cat. no. 55506; Cell Signaling Technology, Inc.), anti-Bcl-2 (1:1,000; cat. no. 60178-1-Ig, ProteinTech Group, Inc.), anti-Bax (1:5,000; cat. no. 60267-1-Ig; ProteinTech Group, Inc.), anti-β-actin (1:1,000; cat. no. 4970; Cell Signaling Technology, Inc.), or anti-GAPDH (1:1,000; cat. no. 2118; Cell Signaling Technology, Inc.).

    Techniques: Cell Culture, Cell Counting, Western Blot, Expressing

    A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of cyclin D1 protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).

    Journal: Cell Death & Disease

    Article Title: Syrosingopine and UK5099 synergistically suppress non-small cell lung cancer by activating the integrated stress response

    doi: 10.1038/s41419-024-06821-4

    Figure Lengend Snippet: A Cell cycle analysis showing the distribution of H661 and PC-9 cells in different cell cycle phases post-treatment with Syrosingopine, UK-5099, or their combination ( n = 3). B Western blot detection of the level of cyclin D1 protein after treatment with Syrosingopine, UK-5099, or their combination ( n = 3). C The effect of Syrosingopine and UK-5099 on the invasion capabilities of H661 and PC-9 cells was determined by Transwell invasion assays ( n = 3). D The effect of Syrosingopine and UK-5099 on the migratory capabilities of H661 and PC-9 cells was determined by a scratch wound assay ( n = 3). E Flow cytometric analysis of apoptosis via Annexin V-FITC/PI staining in H661 and PC-9 cells following exposure to Syrosingopine, UK-5099, or their combination ( n = 3). ( P -values: not significant (ns) p > 0.05, * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001).

    Article Snippet: Antibodies against Cyclin D1 (E3P5S), p-eIF2α (Ser51) (D9G8), eIF2α (D7D3), β-Actin (8H10D10), and Vinculin (E1E9V) were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Cell Cycle Assay, Western Blot, Scratch Wound Assay Assay, Staining