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primary antibody against timp3  (ZenBio)

 
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    ZenBio primary antibody against timp3
    Primary Antibody Against Timp3, supplied by ZenBio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibody against timp3/product/ZenBio
    Average 90 stars, based on 1 article reviews
    primary antibody against timp3 - by Bioz Stars, 2026-03
    90/100 stars

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    primary antibody against timp3  (ZenBio)
    90
    ZenBio primary antibody against timp3
    Primary Antibody Against Timp3, supplied by ZenBio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibody against timp3/product/ZenBio
    Average 90 stars, based on 1 article reviews
    primary antibody against timp3 - by Bioz Stars, 2026-03
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    specic primary antibodies against timp3  (Santa Cruz Biotechnology)
    94
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    Fig. 3 <t>TIMP3</t> was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 30-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. **P < 0.01 or ***P < 0.001 vs. respective control.
    Specic Primary Antibodies Against Timp3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/specic primary antibodies against timp3/product/Santa Cruz Biotechnology
    Average 94 stars, based on 1 article reviews
    specic primary antibodies against timp3 - by Bioz Stars, 2026-03
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    primary antibodies against timp3  (Santa Cruz Biotechnology)
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    Santa Cruz Biotechnology primary antibodies against timp3
    <t>TIMP3</t> was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 3′-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. ** P < 0.01 or *** P < 0.001 vs. respective control.
    Primary Antibodies Against Timp3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against timp3/product/Santa Cruz Biotechnology
    Average 94 stars, based on 1 article reviews
    primary antibodies against timp3 - by Bioz Stars, 2026-03
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    primary antibody against cdkn1b and timp3  (Santa Cruz Biotechnology)
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    <t>TIMP3</t> was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 3′-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. ** P < 0.01 or *** P < 0.001 vs. respective control.
    Primary Antibody Against Cdkn1b And Timp3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibody against cdkn1b and timp3/product/Santa Cruz Biotechnology
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    primary antibodies against timp3 af0265  (Affinity Biologicals)
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    Affinity Biologicals primary antibodies against timp3 af0265
    <t>TIMP3</t> was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 3′-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. ** P < 0.01 or *** P < 0.001 vs. respective control.
    Primary Antibodies Against Timp3 Af0265, supplied by Affinity Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against timp3 af0265/product/Affinity Biologicals
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    primary antibodies against timp3  (Merck KGaA)
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    Merck KGaA primary antibodies against timp3
    <t>TIMP3</t> was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 3′-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. ** P < 0.01 or *** P < 0.001 vs. respective control.
    Primary Antibodies Against Timp3, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against timp3/product/Merck KGaA
    Average 90 stars, based on 1 article reviews
    primary antibodies against timp3 - by Bioz Stars, 2026-03
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    primary antibodies against timp1, timp2, timp3, timp4  (Millipore)
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    Millipore primary antibodies against timp1, timp2, timp3, timp4
    Levels of testicular TIMPs and MPs in response to MEHP exposure by Western blot analysis. Whole-testis homogenates from 28-day-old C57BL/6J mice and total protein from primary Sertoli cell-germ cell cocultures exposed to MEHP (1 g/kg) are examined, and the quantified relative protein levels are shown as column charts. TIMP1, <t>TIMP2,</t> TIMP3, and TIMP4 protein levels (A) and MMP2, MMP9, ADAM10, and ADAM17 protein levels (B) are determined in vivo. After MEHP exposure, TIMP1 and TIMP2 levels are decreased, and MMP2 levels are increased for a short period. C) MMP2 and TIMP2 protein levels in primary coculture cells are analyzed. TIMP2 expression is observed only in primary Sertoli cells and is significantly decreased after MEHP (200 μM) exposure. GC, germ cells; SC, Sertoli cells. ACTB served as the loading control. Asterisks denote significant differences between the treatment and the control (* P < 0.05, Student t-test).
    Primary Antibodies Against Timp1, Timp2, Timp3, Timp4, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against timp1, timp2, timp3, timp4/product/Millipore
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    Fig. 3 TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 30-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. **P < 0.01 or ***P < 0.001 vs. respective control.

    Journal: RSC advances

    Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

    doi: 10.1039/c8ra10552a

    Figure Lengend Snippet: Fig. 3 TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 30-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. **P < 0.01 or ***P < 0.001 vs. respective control.

    Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specic primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or b-actin (sc517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

    Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Construct, Transfection, Quantitative RT-PCR, Western Blot, Control

    Fig. 4 TGF-b1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with different concentration (0, 2, 5 and 10 ng ml1) of TGF- b1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml1 of TGF-b1 at different time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. *P < 0.05 or **P < 0.01 or ***P < 0.001 vs. control.

    Journal: RSC advances

    Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

    doi: 10.1039/c8ra10552a

    Figure Lengend Snippet: Fig. 4 TGF-b1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with different concentration (0, 2, 5 and 10 ng ml1) of TGF- b1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml1 of TGF-b1 at different time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. *P < 0.05 or **P < 0.01 or ***P < 0.001 vs. control.

    Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specic primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or b-actin (sc517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

    Techniques: Expressing, Concentration Assay, Western Blot, Control

    Fig. 5 TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-b1 treatment. HaCaT cells were treated with TGF-b1 (5 ng ml1) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-b1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + pcDNA.

    Journal: RSC advances

    Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

    doi: 10.1039/c8ra10552a

    Figure Lengend Snippet: Fig. 5 TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-b1 treatment. HaCaT cells were treated with TGF-b1 (5 ng ml1) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-b1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + pcDNA.

    Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specic primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or b-actin (sc517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

    Techniques: Migration, Transfection, Quantitative RT-PCR, Expressing, MTT Assay, Transwell Assay, Control

    Fig. 6 MiR-132-mediated pro-proliferation and pro-migration effects were antagonized by TIMP3 in TGF-b1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-b1 (5 ng ml1) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + miR-NC or TGF-b1 + miR-132 + pcDNA.

    Journal: RSC advances

    Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3.

    doi: 10.1039/c8ra10552a

    Figure Lengend Snippet: Fig. 6 MiR-132-mediated pro-proliferation and pro-migration effects were antagonized by TIMP3 in TGF-b1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-b1 (5 ng ml1) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). *P < 0.05 or **P < 0.01 vs. control or TGF-b1 + miR-NC or TGF-b1 + miR-132 + pcDNA.

    Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specic primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or b-actin (sc517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

    Techniques: Migration, Transfection, MTT Assay, Transwell Assay, Control

    TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 3′-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. ** P < 0.01 or *** P < 0.001 vs. respective control.

    Journal: RSC Advances

    Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

    doi: 10.1039/c8ra10552a

    Figure Lengend Snippet: TIMP3 was a direct target of miR-132 in HaCaT cells. (A) The miR-132 binding sites predicted in the 3′-UTR of TIMP3 mRNA and the mutant in seed sites. (B) The relative luciferase activity was detected in HaCaT cells cotransfected with TIMP3-WT or TIMP3-MUT constructs and miR-NC mimics or miR-132 mimics. (C) HaCaT cells were transfected with miR-NC mimics or miR-132 mimics, and the enrichment of TIMP3 mRNA was measured with anti-Ago2 or anti-IgG by qRT-PCR assay. (D) HaCaT cells were transfected with miR-NC mimics, miR-132 mimics, anti-miR-NC or anti-miR-132, followed by the detection of TIMP3 level by Western blot. ** P < 0.01 or *** P < 0.001 vs. respective control.

    Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specific primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or β-actin (sc-517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 °C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

    Techniques: Binding Assay, Mutagenesis, Luciferase, Activity Assay, Construct, Transfection, Quantitative RT-PCR, Western Blot, Control

    TGF-β1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with different concentration (0, 2, 5 and 10 ng ml −1 ) of TGF-β1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml −1 of TGF-β1 at different time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. * P < 0.05 or ** P < 0.01 or *** P < 0.001 vs. control.

    Journal: RSC Advances

    Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

    doi: 10.1039/c8ra10552a

    Figure Lengend Snippet: TGF-β1 repressed the expression of TIMP3. HaCaT (A) and NHEK (B) cells were treated with different concentration (0, 2, 5 and 10 ng ml −1 ) of TGF-β1 for 24 h, and then TIMP3 expression was detected by Western blot. HaCaT (C) and NHEK (D) cells were treated with 5 ng ml −1 of TGF-β1 at different time period (0, 6, 12 and 24 h), followed by the measurement of TIMP3 expression by Western blot. * P < 0.05 or ** P < 0.01 or *** P < 0.001 vs. control.

    Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specific primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or β-actin (sc-517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 °C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

    Techniques: Expressing, Concentration Assay, Western Blot, Control

    TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-β1 treatment. HaCaT cells were treated with TGF-β1 (5 ng ml −1 ) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-β1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. * P < 0.05 or ** P < 0.01 vs. control or TGF-β1 + pcDNA.

    Journal: RSC Advances

    Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

    doi: 10.1039/c8ra10552a

    Figure Lengend Snippet: TIMP3 inhibited the proliferation and migration of HaCaT cells under TGF-β1 treatment. HaCaT cells were treated with TGF-β1 (5 ng ml −1 ) or transfected with pcDNA or pcDNA-TIMP3 prior to TGF-β1 treatment. (A) and (B) qRT-PCR for miR-132 expression in treated cells. (C) and (D) MTT assay for the proliferation ability in treated cells. (E) and (F) Transwell assay for the migration capacity in treated cells. * P < 0.05 or ** P < 0.01 vs. control or TGF-β1 + pcDNA.

    Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specific primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or β-actin (sc-517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 °C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

    Techniques: Migration, Transfection, Quantitative RT-PCR, Expressing, MTT Assay, Transwell Assay, Control

    MiR-132-mediated pro-proliferation and pro-migration effects were antagonized by TIMP3 in TGF-β1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-β1 (5 ng ml −1 ) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). * P < 0.05 or ** P < 0.01 vs. control or TGF-β1 + miR-NC or TGF-β1 + miR-132 + pcDNA.

    Journal: RSC Advances

    Article Title: Retracted Article: MiR-132 enhances proliferation and migration of HaCaT cells by targeting TIMP3

    doi: 10.1039/c8ra10552a

    Figure Lengend Snippet: MiR-132-mediated pro-proliferation and pro-migration effects were antagonized by TIMP3 in TGF-β1-treated HaCaT cells. HaCaT cells were transfected with miR-132 mimics + pcDNA, miR-132 mimics + pcDNA-TIMP3, anti-miR-132 + si-NC or anti-miR-132 + si-TIMP3 prior to TGF-β1 (5 ng ml −1 ) treatment, followed by the detection of cell proliferation ability by MTT assay (A) and (B), cell migration capacity by transwell assay (C) and (D). * P < 0.05 or ** P < 0.01 vs. control or TGF-β1 + miR-NC or TGF-β1 + miR-132 + pcDNA.

    Article Snippet: Blocked by a buffer containing 5% non-fat milk in TBS with 0.1% Tween-20, the membrances were probed overnight with specific primary antibodies against TIMP3 (sc-373839, Santa Cruz Biotechnology, Santa Cruz, CA, USA; dilution 1 : 500) or β-actin (sc-517582, Santa Cruz Biotechnology; dilution 1 : 1000) at 4 °C, followed by the incubation with HRP-conjugated secondary antibodies (sc-516102, Santa Cruz Biotechnology; dilution 1 : 5000).

    Techniques: Migration, Transfection, MTT Assay, Transwell Assay, Control

    Levels of testicular TIMPs and MPs in response to MEHP exposure by Western blot analysis. Whole-testis homogenates from 28-day-old C57BL/6J mice and total protein from primary Sertoli cell-germ cell cocultures exposed to MEHP (1 g/kg) are examined, and the quantified relative protein levels are shown as column charts. TIMP1, TIMP2, TIMP3, and TIMP4 protein levels (A) and MMP2, MMP9, ADAM10, and ADAM17 protein levels (B) are determined in vivo. After MEHP exposure, TIMP1 and TIMP2 levels are decreased, and MMP2 levels are increased for a short period. C) MMP2 and TIMP2 protein levels in primary coculture cells are analyzed. TIMP2 expression is observed only in primary Sertoli cells and is significantly decreased after MEHP (200 μM) exposure. GC, germ cells; SC, Sertoli cells. ACTB served as the loading control. Asterisks denote significant differences between the treatment and the control (* P < 0.05, Student t-test).

    Journal: Biology of Reproduction

    Article Title: TNF Alpha-Mediated Disruption of Spermatogenesis in Response to Sertoli Cell Injury in Rodents Is Partially Regulated by MMP2 1

    doi: 10.1095/biolreprod.108.073122

    Figure Lengend Snippet: Levels of testicular TIMPs and MPs in response to MEHP exposure by Western blot analysis. Whole-testis homogenates from 28-day-old C57BL/6J mice and total protein from primary Sertoli cell-germ cell cocultures exposed to MEHP (1 g/kg) are examined, and the quantified relative protein levels are shown as column charts. TIMP1, TIMP2, TIMP3, and TIMP4 protein levels (A) and MMP2, MMP9, ADAM10, and ADAM17 protein levels (B) are determined in vivo. After MEHP exposure, TIMP1 and TIMP2 levels are decreased, and MMP2 levels are increased for a short period. C) MMP2 and TIMP2 protein levels in primary coculture cells are analyzed. TIMP2 expression is observed only in primary Sertoli cells and is significantly decreased after MEHP (200 μM) exposure. GC, germ cells; SC, Sertoli cells. ACTB served as the loading control. Asterisks denote significant differences between the treatment and the control (* P < 0.05, Student t-test).

    Article Snippet: Total cellular proteins (30 μg) were tested using primary antibodies against TIMP1, TIMP2, TIMP3, and TIMP4 (1:1000; Chemicon, Temecula, CA); matrix MP 2 (MMP2) and MMP9 (1:1000; Abcam Inc., Cambridge, MA); FASL (1:500; Santa Cruz Biotechnology Inc., Santa Cruz, CA); TNFA (1:500; R&D Systems Inc., Minneapolis, MN); a distintegrin and MP 10 (ADAM10) (1:500; R&D Systems Inc.), ADAM17 (1:500; Abcam Inc.); and β-actin (ACTB) (1:500; Santa Cruz Biotechnology Inc.) coupled with horseradish peroxidase-conjugated secondary antibodies (1:5000; Santa Cruz Biotechnology Inc.).

    Techniques: Western Blot, In Vivo, Expressing

    A proposed working model of the complex Sertoli cell-germ cell interaction describing the mechanism by which Sertoli cells trigger germ cell apoptosis following MEHP-induced Sertoli cell injury. First, MEHP-induced decreases in TIMP2 expression in Sertoli cells allow for the increased activation of MMP2. Active MMP2 can then cleave membrane-bound TNFA produced by germ cells to generate the soluble active form of TNFA. The binding of sTNFA with Sertoli cell TNFRSF1A triggers the NFKB1 signaling pathway and robustly induces FASL expression. An increase in Sertoli cell FASL allows for the instigation of germ cell apoptosis by the classic death receptor pathway.

    Journal: Biology of Reproduction

    Article Title: TNF Alpha-Mediated Disruption of Spermatogenesis in Response to Sertoli Cell Injury in Rodents Is Partially Regulated by MMP2 1

    doi: 10.1095/biolreprod.108.073122

    Figure Lengend Snippet: A proposed working model of the complex Sertoli cell-germ cell interaction describing the mechanism by which Sertoli cells trigger germ cell apoptosis following MEHP-induced Sertoli cell injury. First, MEHP-induced decreases in TIMP2 expression in Sertoli cells allow for the increased activation of MMP2. Active MMP2 can then cleave membrane-bound TNFA produced by germ cells to generate the soluble active form of TNFA. The binding of sTNFA with Sertoli cell TNFRSF1A triggers the NFKB1 signaling pathway and robustly induces FASL expression. An increase in Sertoli cell FASL allows for the instigation of germ cell apoptosis by the classic death receptor pathway.

    Article Snippet: Total cellular proteins (30 μg) were tested using primary antibodies against TIMP1, TIMP2, TIMP3, and TIMP4 (1:1000; Chemicon, Temecula, CA); matrix MP 2 (MMP2) and MMP9 (1:1000; Abcam Inc., Cambridge, MA); FASL (1:500; Santa Cruz Biotechnology Inc., Santa Cruz, CA); TNFA (1:500; R&D Systems Inc., Minneapolis, MN); a distintegrin and MP 10 (ADAM10) (1:500; R&D Systems Inc.), ADAM17 (1:500; Abcam Inc.); and β-actin (ACTB) (1:500; Santa Cruz Biotechnology Inc.) coupled with horseradish peroxidase-conjugated secondary antibodies (1:5000; Santa Cruz Biotechnology Inc.).

    Techniques: Expressing, Activation Assay, Produced, Binding Assay