cyclin d2 Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Thermo Fisher anti cyclin d2
    DHPS inhibition attenuates harmine-induced β cell proliferation in mouse and human islets., ( A ) Islets were isolated from male 8–9 week old CD1 mice, then treated in vitro with vehicle (V), harmine (H), and Gc7 (G). Representative immunoblot (top) and quantification of protein levels (bottom) for <t>Cyclin-D2.</t> Data are from N=6 mice per group. ( B ) Islets were isolated from male 8–9 week old CD1 mice, then treated in vitro with vehicle (V), harmine (H), and Gc7, followed by flow cytometry analysis of dispersed islet cells immunostained for phospho-histone H3. Data are from N=4–6 mice per group. ( C ). Islets were isolated from control (Cre+) and Dhps Δβ (βKO) mice, then treated in vitro with vehicle (V) or harmine (H), then percent of β cells that immunostained for Ki67 was calculated. Data are from N=3 animals per group. ( D ) Human islets from 4 donors were treated in vitro with vehicle, harmine, and/or Gc7 (G), dispersed, immunostained for phospho-histone H3, then subjected to flow cytometry analysis. Graphs show a single technical replicate for each donor. Data in A, B, and C are presented as mean ± SEM; * p -value
    Anti Cyclin D2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cyclin d2/product/Thermo Fisher
    Average 94 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    anti cyclin d2 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    93
    Santa Cruz Biotechnology cyclin d2
    <t>Cyclin</t> D2 deficiency reduces proliferation following Apc loss
    Cyclin D2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 815 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cyclin d2/product/Santa Cruz Biotechnology
    Average 93 stars, based on 815 article reviews
    Price from $9.99 to $1999.99
    cyclin d2 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    92
    Cell Signaling Technology Inc cyclin d2
    (A-C), in situ hybridisation for miR-133a-3p expression in normal (C) and diseased (A and B) retinas. (D), luciferase assay to study the correlation of miR-133a-3p with <t>cyclin</t> D2 expression in 661 photoreceptor cells. The expression of miR-133a-3p in the outer nuclear layer (ONL) was increased after induced Müller cell ablation. Co-transfection of 661w cells using a vector containing 3’UTR region of the cyclin D2 gene along with a miR-133a-3p mimic (Vector + miR) induced a significant increase in the normalised firefly luciferase activity compared with cells co-transfected using the vector with a control miRNA (Vector + Ctrl) (*P
    Cyclin D2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 609 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cyclin d2/product/Cell Signaling Technology Inc
    Average 92 stars, based on 609 article reviews
    Price from $9.99 to $1999.99
    cyclin d2 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    93
    Abcam cyclin d2
    Nox4 overexpression activates c-myc resulting in increased <t>cyclin</t> D2 expression. (A) Representative immunoblot showing increased c-myc phosphorylation in 2 week-old Nox4 Tg hearts compared with Wt littermate controls. The histogram represents the ratio of phosphorylated c-myc to total c-myc protein levels (n = 3). (B) Representative immunoblot and quantitative histogram showing phosphorylated and total c-myc levels in NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence or absence of PD98059 as indicated (n = 3). (C) Q-PCR analyses of c-myc mRNA expression, relative to that of β-actin (n = 3), and (D) representative immunoblot indicating c-myc protein expression, in NRCs after 48 h of c-myc siRNA treatment as indicated. β-Actin is shown as a loading control for protein expression. (E) Representative immunoblot and quantitative histogram showing cyclin D2 protein expression, relative to that of β-actin, in NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence of scrambled or c-myc siRNA as indicated (n = 3). All data are presented as mean ± S.E. * P
    Cyclin D2, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 764 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cyclin d2/product/Abcam
    Average 93 stars, based on 764 article reviews
    Price from $9.99 to $1999.99
    cyclin d2 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    90
    Santa Cruz Biotechnology anti cyclin d2
    The expression of D-cyclins in mantle cell lymphoma The differential expression of <t>D-cyclin</t> staining in two case examples of mantle cell lymphoma is shown. Case 1 shows intense cyclin D1 and weak <t>cyclin</t> D2 staining. Case 2 shows weak cyclin D1 and strong cyclin D2 staining. Both cases lack staining for cyclin D3.
    Anti Cyclin D2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cyclin d2/product/Santa Cruz Biotechnology
    Average 90 stars, based on 258 article reviews
    Price from $9.99 to $1999.99
    anti cyclin d2 - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    88
    Santa Cruz Biotechnology rabbit anti cyclin d2
    <t>Cyclin</t> D2 is enriched in the ventral peripheral retina during the temporal window of ipsilateral RGC genesis. IHC in coronal cryosections of E11.5 ( A ), E12.5 ( B ), E14.5 ( C ), E15.5 ( D ), and E16.5 ( E ) retina. Cyclin D2 is expressed in the lens and the retinal marginal zone bordering RGCs (labeled with Brn3). Cyclin D2 is highly expressed in the ventral periphery compared with the dorsal periphery within neural retina (white arrows). The cyclin D2 and Brn3 domains are separated by a large gap at E11.5 ( A ). At E13.5 ( B ) and E14.5 ( C ), cyclin D2 + cells intermingle with Brn3 + RGCs only in ventral retina (yellow arrows point to boundary) and have sharp boundaries in dorsal retina. At E16.5 ( E ), cyclin D2 expression is reduced in the retinal periphery and is no longer asymmetric in dorsal and ventral retinal (white arrows). Scale bars, 100 μm.
    Rabbit Anti Cyclin D2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti cyclin d2/product/Santa Cruz Biotechnology
    Average 88 stars, based on 65 article reviews
    Price from $9.99 to $1999.99
    rabbit anti cyclin d2 - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

    92
    Thermo Fisher cyclin d2
    Effect of AMPK activation on <t>cyclin</t> D2 mRNA expression in granulosa cells. Granulosa cells from 3-d estradiol primed immature rats were cultured in serum-free, phenol red-free medium overnight followed by incubation with AMPK activator (AICAR) for 1 h.
    Cyclin D2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 175 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cyclin d2/product/Thermo Fisher
    Average 92 stars, based on 175 article reviews
    Price from $9.99 to $1999.99
    cyclin d2 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc anti cyclin d2
    <t>Cyclin</t> D/CDK4 negatively regulates PD-L1 protein stability a, b, Immunoblot (IB) analysis of whole cell lysates (WCL) derived from wild type (WT), cyclin A1 −/− A2 −/− or WT, cyclin E1 −/− E2 −/− MEFs. c, Quantitative real-time PCR (qRT-PCR) analysis of relative mRNA levels of PD-L1 from wild type MEFs and cyclin D1 −/− D2 −/− D3 −/− MEFs. Data were represented as mean ± s.d, n = 5. d, Cell cycle profiles for WT and cyclin D1 −/− D2 −/− D3 −/− MEFs, which were labeled with BrdU and analyzed by FACS. e, IB analysis of WCL derived from cyclin D1 fl/fl D2 −/− D3 fl/fl MEFs with or without depleting cyclin D1 and cyclin D3 by pLenti-Cre via viral infection (pLenti-EGFP as a negative control), selected with puromycin (1 μg/ml) for 72 hours before harvesting. f, IB analysis of WCL derived from cyclin D1 −/− D2 −/− D3 −/− MEFs stably reintroducing cyclin D1 , <t>cyclin</t> D2 , or cyclin D3 , respectively, with empty vector (EV) as a negative control. g, IB analysis of WCL derived from mouse mammary tumors induced by MMTV- c-Myc with/without genetic depletion of cyclin D1 . n = 5 mice per experimental group. h, IB analysis of WCL derived from WCL derived from wild type and cdk6 −/− MEFs. i, j, IB analysis of WCL derived from MDA-MB-231 cells stably expressing shCDK6 or shCDK2 as well as shScr as a negative control, respectively. k, l, IB analysis of WCL derived from MDA-MB-231 cells transfected with indicated constructs (k) and the intensity of PD-L1 band was quantified by the ImageJ software (l) . m, IB analysis of WCL derived from MDA-MB-231 cells depleted of Rb (with shScr as a negative control) treated with the CDK4/6 inhibitor, palbociclib, where indicated. n, o, IB analysis of WCL derived from mouse CT26 or 4T1 tumor cell lines treated with or without the CDK4/6 inhibitor, palbociclib or ribociclib, respectively. p, q, IB analysis of WCL derived from MDA-MB-231 cells pre-treated with palbociclib (1 μM) for 36 hours before treatment with cycloheximide (CHX) for the indicated time points (p) and PD-L1 protein abundance was quantified by the ImageJ and plotted as indicated (q) . r, IB analysis of WCL derived from 19 different cancer cell lines with indicated antibodies. s–u, IB analysis of WCL derived from MCF7, T47D or HLF stably expressing p16 as well as EV as a negative control. v–x, IB analysis of WCL derived from MDA-MB-436, BT549 or HCC1937 stably expressing three independent shRNAs against p16 as well as shScr as a negative control.
    Anti Cyclin D2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cyclin d2/product/Cell Signaling Technology Inc
    Average 94 stars, based on 52 article reviews
    Price from $9.99 to $1999.99
    anti cyclin d2 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    89
    SLIT2 LTD cyclind2
    <t>Cyclin</t> D/CDK4 negatively regulates PD-L1 protein stability a, b, Immunoblot (IB) analysis of whole cell lysates (WCL) derived from wild type (WT), cyclin A1 −/− A2 −/− or WT, cyclin E1 −/− E2 −/− MEFs. c, Quantitative real-time PCR (qRT-PCR) analysis of relative mRNA levels of PD-L1 from wild type MEFs and cyclin D1 −/− D2 −/− D3 −/− MEFs. Data were represented as mean ± s.d, n = 5. d, Cell cycle profiles for WT and cyclin D1 −/− D2 −/− D3 −/− MEFs, which were labeled with BrdU and analyzed by FACS. e, IB analysis of WCL derived from cyclin D1 fl/fl D2 −/− D3 fl/fl MEFs with or without depleting cyclin D1 and cyclin D3 by pLenti-Cre via viral infection (pLenti-EGFP as a negative control), selected with puromycin (1 μg/ml) for 72 hours before harvesting. f, IB analysis of WCL derived from cyclin D1 −/− D2 −/− D3 −/− MEFs stably reintroducing cyclin D1 , <t>cyclin</t> D2 , or cyclin D3 , respectively, with empty vector (EV) as a negative control. g, IB analysis of WCL derived from mouse mammary tumors induced by MMTV- c-Myc with/without genetic depletion of cyclin D1 . n = 5 mice per experimental group. h, IB analysis of WCL derived from WCL derived from wild type and cdk6 −/− MEFs. i, j, IB analysis of WCL derived from MDA-MB-231 cells stably expressing shCDK6 or shCDK2 as well as shScr as a negative control, respectively. k, l, IB analysis of WCL derived from MDA-MB-231 cells transfected with indicated constructs (k) and the intensity of PD-L1 band was quantified by the ImageJ software (l) . m, IB analysis of WCL derived from MDA-MB-231 cells depleted of Rb (with shScr as a negative control) treated with the CDK4/6 inhibitor, palbociclib, where indicated. n, o, IB analysis of WCL derived from mouse CT26 or 4T1 tumor cell lines treated with or without the CDK4/6 inhibitor, palbociclib or ribociclib, respectively. p, q, IB analysis of WCL derived from MDA-MB-231 cells pre-treated with palbociclib (1 μM) for 36 hours before treatment with cycloheximide (CHX) for the indicated time points (p) and PD-L1 protein abundance was quantified by the ImageJ and plotted as indicated (q) . r, IB analysis of WCL derived from 19 different cancer cell lines with indicated antibodies. s–u, IB analysis of WCL derived from MCF7, T47D or HLF stably expressing p16 as well as EV as a negative control. v–x, IB analysis of WCL derived from MDA-MB-436, BT549 or HCC1937 stably expressing three independent shRNAs against p16 as well as shScr as a negative control.
    Cyclind2, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 89/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cyclind2/product/SLIT2 LTD
    Average 89 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
    cyclind2 - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    95
    Abcam anti cyclin d2 antibody epr19659
    <t>Cyclin</t> D/CDK4 negatively regulates PD-L1 protein stability a, b, Immunoblot (IB) analysis of whole cell lysates (WCL) derived from wild type (WT), cyclin A1 −/− A2 −/− or WT, cyclin E1 −/− E2 −/− MEFs. c, Quantitative real-time PCR (qRT-PCR) analysis of relative mRNA levels of PD-L1 from wild type MEFs and cyclin D1 −/− D2 −/− D3 −/− MEFs. Data were represented as mean ± s.d, n = 5. d, Cell cycle profiles for WT and cyclin D1 −/− D2 −/− D3 −/− MEFs, which were labeled with BrdU and analyzed by FACS. e, IB analysis of WCL derived from cyclin D1 fl/fl D2 −/− D3 fl/fl MEFs with or without depleting cyclin D1 and cyclin D3 by pLenti-Cre via viral infection (pLenti-EGFP as a negative control), selected with puromycin (1 μg/ml) for 72 hours before harvesting. f, IB analysis of WCL derived from cyclin D1 −/− D2 −/− D3 −/− MEFs stably reintroducing cyclin D1 , <t>cyclin</t> D2 , or cyclin D3 , respectively, with empty vector (EV) as a negative control. g, IB analysis of WCL derived from mouse mammary tumors induced by MMTV- c-Myc with/without genetic depletion of cyclin D1 . n = 5 mice per experimental group. h, IB analysis of WCL derived from WCL derived from wild type and cdk6 −/− MEFs. i, j, IB analysis of WCL derived from MDA-MB-231 cells stably expressing shCDK6 or shCDK2 as well as shScr as a negative control, respectively. k, l, IB analysis of WCL derived from MDA-MB-231 cells transfected with indicated constructs (k) and the intensity of PD-L1 band was quantified by the ImageJ software (l) . m, IB analysis of WCL derived from MDA-MB-231 cells depleted of Rb (with shScr as a negative control) treated with the CDK4/6 inhibitor, palbociclib, where indicated. n, o, IB analysis of WCL derived from mouse CT26 or 4T1 tumor cell lines treated with or without the CDK4/6 inhibitor, palbociclib or ribociclib, respectively. p, q, IB analysis of WCL derived from MDA-MB-231 cells pre-treated with palbociclib (1 μM) for 36 hours before treatment with cycloheximide (CHX) for the indicated time points (p) and PD-L1 protein abundance was quantified by the ImageJ and plotted as indicated (q) . r, IB analysis of WCL derived from 19 different cancer cell lines with indicated antibodies. s–u, IB analysis of WCL derived from MCF7, T47D or HLF stably expressing p16 as well as EV as a negative control. v–x, IB analysis of WCL derived from MDA-MB-436, BT549 or HCC1937 stably expressing three independent shRNAs against p16 as well as shScr as a negative control.
    Anti Cyclin D2 Antibody Epr19659, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cyclin d2 antibody epr19659/product/Abcam
    Average 95 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    anti cyclin d2 antibody epr19659 - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    94
    Abcam anti cyclin d2
    5-AZA blocked dexamethasone-induced downregulation of <t>cyclin</t> D2 protein and mRNA abundance in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and treated with dexamethasone (DEX) in the absence or presence of 5-aza-2'-deoxycytidine (5-AZA) for 48 hours. Cyclin D2 protein and mRNA abundance was determined by Western blot and quantitative real-time PCR, respectively. Data are mean ± S.E.M., n = 4 or 5, from different experiments. * P
    Anti Cyclin D2, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cyclin d2/product/Abcam
    Average 94 stars, based on 38 article reviews
    Price from $9.99 to $1999.99
    anti cyclin d2 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    90
    Becton Dickinson anti cyclin d2
    Knockdown of CITED1 upregulates the expression of the CKIs p21 and p27. a Western blotting analysis was performed to detect the expression levels of the cell cycle regulators <t>cyclin</t> A2, cyclin B1, cyclin D1, cyclin D2, cyclin D3, cyclin E1, cyclin E2, CDK2, CDK4, CDK6, p21 Cip1 , and p27 Kip1 in the indicated cells. α-tubulin was used as the loading control. b Silencing of CITED1 in the studied cells significantly inhibited the phosphorylation of pRb at the Ser608 residue. α-tubulin served as the sample loading control. c Silencing of CITED1 attenuated E2F transcriptional activity according to the E2F-luc reporter assay. The results are derived from three independent experiments and are expressed as the mean ± SD. *P
    Anti Cyclin D2, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cyclin d2/product/Becton Dickinson
    Average 90 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    anti cyclin d2 - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    92
    rdi research diagnostics cyclin d2
    Pilomatricomas express cyclins D1, D2, and D3. A: H E stain of pilomatricoma. Arrow indicates shadow cells; arrowhead indicates basoloid cells (×200), Tumor cells are focally positive for <t>cyclin</t> D1 ( B ), <t>cyclin</t> D2 ( C ), and cyclin D3 ( D ) ( arrows , ×200).
    Cyclin D2, supplied by rdi research diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cyclin d2/product/rdi research diagnostics
    Average 92 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    cyclin d2 - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    93
    R&D Systems human mouse cyclin d1 d2 cross reactive antibody
    Pilomatricomas express cyclins D1, D2, and D3. A: H E stain of pilomatricoma. Arrow indicates shadow cells; arrowhead indicates basoloid cells (×200), Tumor cells are focally positive for <t>cyclin</t> D1 ( B ), <t>cyclin</t> D2 ( C ), and cyclin D3 ( D ) ( arrows , ×200).
    Human Mouse Cyclin D1 D2 Cross Reactive Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human mouse cyclin d1 d2 cross reactive antibody/product/R&D Systems
    Average 93 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    human mouse cyclin d1 d2 cross reactive antibody - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    90
    Santa Cruz Biotechnology rabbit polyclonal anti cyclin d2
    AZD1480 reduces the tumor growth of Kms.11 xenograft in mice associated with inhibition of JAK2/STAT3 and FGFR3 signaling and <t>Cyclin</t> D2 in vivo
    Rabbit Polyclonal Anti Cyclin D2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti cyclin d2/product/Santa Cruz Biotechnology
    Average 90 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti cyclin d2 - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    85
    Abcam mouse monoclonal cyclin d2
    RNAi-mediated knockdown of <t>cyclin</t> D3 induces G 2 /M arrest and polyploidy. (A) MLE cells were transfected with either scrambled RNA or <t>cyclin</t> D2 siRNA or cyclin D3 siRNA. 72 h later, cells were harvested; lysates were resolved on SDS-PAGE prior to cyclin
    Mouse Monoclonal Cyclin D2, supplied by Abcam, used in various techniques. Bioz Stars score: 85/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal cyclin d2/product/Abcam
    Average 85 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal cyclin d2 - by Bioz Stars, 2020-09
    85/100 stars
      Buy from Supplier

    Image Search Results


    DHPS inhibition attenuates harmine-induced β cell proliferation in mouse and human islets., ( A ) Islets were isolated from male 8–9 week old CD1 mice, then treated in vitro with vehicle (V), harmine (H), and Gc7 (G). Representative immunoblot (top) and quantification of protein levels (bottom) for Cyclin-D2. Data are from N=6 mice per group. ( B ) Islets were isolated from male 8–9 week old CD1 mice, then treated in vitro with vehicle (V), harmine (H), and Gc7, followed by flow cytometry analysis of dispersed islet cells immunostained for phospho-histone H3. Data are from N=4–6 mice per group. ( C ). Islets were isolated from control (Cre+) and Dhps Δβ (βKO) mice, then treated in vitro with vehicle (V) or harmine (H), then percent of β cells that immunostained for Ki67 was calculated. Data are from N=3 animals per group. ( D ) Human islets from 4 donors were treated in vitro with vehicle, harmine, and/or Gc7 (G), dispersed, immunostained for phospho-histone H3, then subjected to flow cytometry analysis. Graphs show a single technical replicate for each donor. Data in A, B, and C are presented as mean ± SEM; * p -value

    Journal: Science signaling

    Article Title: Hypusine biosynthesis in β cells links polyamine metabolism to facultative cellular proliferation to maintain glucose homeostasis

    doi: 10.1126/scisignal.aax0715

    Figure Lengend Snippet: DHPS inhibition attenuates harmine-induced β cell proliferation in mouse and human islets., ( A ) Islets were isolated from male 8–9 week old CD1 mice, then treated in vitro with vehicle (V), harmine (H), and Gc7 (G). Representative immunoblot (top) and quantification of protein levels (bottom) for Cyclin-D2. Data are from N=6 mice per group. ( B ) Islets were isolated from male 8–9 week old CD1 mice, then treated in vitro with vehicle (V), harmine (H), and Gc7, followed by flow cytometry analysis of dispersed islet cells immunostained for phospho-histone H3. Data are from N=4–6 mice per group. ( C ). Islets were isolated from control (Cre+) and Dhps Δβ (βKO) mice, then treated in vitro with vehicle (V) or harmine (H), then percent of β cells that immunostained for Ki67 was calculated. Data are from N=3 animals per group. ( D ) Human islets from 4 donors were treated in vitro with vehicle, harmine, and/or Gc7 (G), dispersed, immunostained for phospho-histone H3, then subjected to flow cytometry analysis. Graphs show a single technical replicate for each donor. Data in A, B, and C are presented as mean ± SEM; * p -value

    Article Snippet: For immunoblots, antibodies were as follows: anti-eIF5AHyp (1:3000) ( ); anti-eIF5ATotal (1:3000, Clone 26, BD Biosciences); anti-ERK1/2 (1:1,000, sc-94, Santa Cruz); anti-Cyclin-D2 (1:1,000, AB-2, Thermo Fisher Scientific); anti-Cyclin-D1 (1:1000, AB-1, Thermo Fisher Scientific); anti-β-tubulin (1:3000, 9F3, Cell Signaling); anti-DHPS (1:1000, sc-365077, Santa Cruz).

    Techniques: Inhibition, Isolation, Mouse Assay, In Vitro, Flow Cytometry

    Decreased mRNA translation of Cyclin-D2 in Dhps Δβ mice. ( A ) Schematic showing experimental design. Control (Cre+) and Dhps Δβ (βKO) mice were fed a HFD for 1 week and islets were isolated and subjected to RT-PCR and polyribosome profiling (PRP) analysis. ( B ) Quantitative RT-PCR from islets for the genes indicated. Data from N=5–6 animals per group. ( C ) Representative immunoblot analysis from islets for Cyclin-D2, Cyclin-D1, and Erk½ (left) and quantification of immunoblots (right). Data from N=3–5 animals per group. ( D ) PRP of isolated islets (left) and quantification of polyribosome:monoribosome ratio (right). Data from N=3 mice per group. ( E ) Quantitative RT-PCR of Cycd2 in polyribosomal fractions. Data from N=3–4 mice per group. ( F ) Quantitative RT-PCR of Cycd1 in polyribosomal fractions. Data from N=3–4 mice per group. Data presented as mean ± SEM; * p -value

    Journal: Science signaling

    Article Title: Hypusine biosynthesis in β cells links polyamine metabolism to facultative cellular proliferation to maintain glucose homeostasis

    doi: 10.1126/scisignal.aax0715

    Figure Lengend Snippet: Decreased mRNA translation of Cyclin-D2 in Dhps Δβ mice. ( A ) Schematic showing experimental design. Control (Cre+) and Dhps Δβ (βKO) mice were fed a HFD for 1 week and islets were isolated and subjected to RT-PCR and polyribosome profiling (PRP) analysis. ( B ) Quantitative RT-PCR from islets for the genes indicated. Data from N=5–6 animals per group. ( C ) Representative immunoblot analysis from islets for Cyclin-D2, Cyclin-D1, and Erk½ (left) and quantification of immunoblots (right). Data from N=3–5 animals per group. ( D ) PRP of isolated islets (left) and quantification of polyribosome:monoribosome ratio (right). Data from N=3 mice per group. ( E ) Quantitative RT-PCR of Cycd2 in polyribosomal fractions. Data from N=3–4 mice per group. ( F ) Quantitative RT-PCR of Cycd1 in polyribosomal fractions. Data from N=3–4 mice per group. Data presented as mean ± SEM; * p -value

    Article Snippet: For immunoblots, antibodies were as follows: anti-eIF5AHyp (1:3000) ( ); anti-eIF5ATotal (1:3000, Clone 26, BD Biosciences); anti-ERK1/2 (1:1,000, sc-94, Santa Cruz); anti-Cyclin-D2 (1:1,000, AB-2, Thermo Fisher Scientific); anti-Cyclin-D1 (1:1000, AB-1, Thermo Fisher Scientific); anti-β-tubulin (1:3000, 9F3, Cell Signaling); anti-DHPS (1:1000, sc-365077, Santa Cruz).

    Techniques: Mouse Assay, Isolation, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

    Proposed pathway linking PKCζ, c-Myc, and polyamines to adaptive proliferation in β cells. The schematic diagram shows the proposed positioning of the PI3K- PKCζ-mTOR pathway (left arm of the diagram) and the polyamine-eIF5A pathway (right arm of the diagram) relative to the adaptive translational and proliferative responses. The hierarchy of the factors depicted in red (PKCζ, c-Myc, ODC, and DHPS) have been shown in this study or others to serve as key nodes through which proliferative responses are mediated – deletion or inhibition of these factors severely restricts the ability of the β cell to produce Cyclin-D2 and undergo replication. This study demonstrated that DHPS is downstream of PKCζ, c-Myc, and ODC functions. This study does not explicitly rule out the possibility that PKCζ might directly or indirectly regulate the function of DHPS (shown as a dashed line with a question mark).

    Journal: Science signaling

    Article Title: Hypusine biosynthesis in β cells links polyamine metabolism to facultative cellular proliferation to maintain glucose homeostasis

    doi: 10.1126/scisignal.aax0715

    Figure Lengend Snippet: Proposed pathway linking PKCζ, c-Myc, and polyamines to adaptive proliferation in β cells. The schematic diagram shows the proposed positioning of the PI3K- PKCζ-mTOR pathway (left arm of the diagram) and the polyamine-eIF5A pathway (right arm of the diagram) relative to the adaptive translational and proliferative responses. The hierarchy of the factors depicted in red (PKCζ, c-Myc, ODC, and DHPS) have been shown in this study or others to serve as key nodes through which proliferative responses are mediated – deletion or inhibition of these factors severely restricts the ability of the β cell to produce Cyclin-D2 and undergo replication. This study demonstrated that DHPS is downstream of PKCζ, c-Myc, and ODC functions. This study does not explicitly rule out the possibility that PKCζ might directly or indirectly regulate the function of DHPS (shown as a dashed line with a question mark).

    Article Snippet: For immunoblots, antibodies were as follows: anti-eIF5AHyp (1:3000) ( ); anti-eIF5ATotal (1:3000, Clone 26, BD Biosciences); anti-ERK1/2 (1:1,000, sc-94, Santa Cruz); anti-Cyclin-D2 (1:1,000, AB-2, Thermo Fisher Scientific); anti-Cyclin-D1 (1:1000, AB-1, Thermo Fisher Scientific); anti-β-tubulin (1:3000, 9F3, Cell Signaling); anti-DHPS (1:1000, sc-365077, Santa Cruz).

    Techniques: Inhibition

    Cyclin D2 deficiency reduces proliferation following Apc loss

    Journal: Cancer research

    Article Title: Cyclin D2-CDK4/6 is required for efficient proliferation and tumorigenesis following Apc loss

    doi: 10.1158/0008-5472.CAN-10-0315

    Figure Lengend Snippet: Cyclin D2 deficiency reduces proliferation following Apc loss

    Article Snippet: In this study we have shown that Cyclin D2 is required for efficient proliferation and tumorigenesis following Apc loss.

    Techniques:

    Cyclin D2 deficiency reduces tumourigenesis in the Apc Min /+ mouse

    Journal: Cancer research

    Article Title: Cyclin D2-CDK4/6 is required for efficient proliferation and tumorigenesis following Apc loss

    doi: 10.1158/0008-5472.CAN-10-0315

    Figure Lengend Snippet: Cyclin D2 deficiency reduces tumourigenesis in the Apc Min /+ mouse

    Article Snippet: In this study we have shown that Cyclin D2 is required for efficient proliferation and tumorigenesis following Apc loss.

    Techniques:

    Cyclin D2 deficiency reduces proliferation rates in adenomas from the Apc Min /+ mouse

    Journal: Cancer research

    Article Title: Cyclin D2-CDK4/6 is required for efficient proliferation and tumorigenesis following Apc loss

    doi: 10.1158/0008-5472.CAN-10-0315

    Figure Lengend Snippet: Cyclin D2 deficiency reduces proliferation rates in adenomas from the Apc Min /+ mouse

    Article Snippet: In this study we have shown that Cyclin D2 is required for efficient proliferation and tumorigenesis following Apc loss.

    Techniques:

    Cyclin D1 haploinsufficiency further suppresses tumorigenesis and tumor proliferation when combined with Cyclin D2 deficiency

    Journal: Cancer research

    Article Title: Cyclin D2-CDK4/6 is required for efficient proliferation and tumorigenesis following Apc loss

    doi: 10.1158/0008-5472.CAN-10-0315

    Figure Lengend Snippet: Cyclin D1 haploinsufficiency further suppresses tumorigenesis and tumor proliferation when combined with Cyclin D2 deficiency

    Article Snippet: In this study we have shown that Cyclin D2 is required for efficient proliferation and tumorigenesis following Apc loss.

    Techniques:

    Cyclin D2 is upregulated immediately following loss of Apc and is c-Myc dependent

    Journal: Cancer research

    Article Title: Cyclin D2-CDK4/6 is required for efficient proliferation and tumorigenesis following Apc loss

    doi: 10.1158/0008-5472.CAN-10-0315

    Figure Lengend Snippet: Cyclin D2 is upregulated immediately following loss of Apc and is c-Myc dependent

    Article Snippet: In this study we have shown that Cyclin D2 is required for efficient proliferation and tumorigenesis following Apc loss.

    Techniques:

    Translational control during endothelial differentiation and rapamycin treatment . a) Preparation of total RNA and ribosome-enriched RNA. RoSH2 cells, RoSH2 induced to differentiate by plating on matrigel for 48 hours and RoSH2 cells treated with 50 ηM rapamycin for 48 hours were harvested. Total and ribosome-enriched RNAs were quantitated by absorbance at 260 nm. 2 μg of RNA from each of the two cellular fractions were separated on a 1.0% agarose gel, stained with ethidium bromide and visualized under UV illumination. Top panel Representative RNA samples from total and ribosome-enriched RNAs prepared from undifferentiated (Undiff), differentiated (Diff) and rapamycin-treated (Rapa) RoSH2 cells are shown. Bottom panel For each sample, 1 μl of serially diluted RNA sample mixed with 1 μl of 0.5 ηg/μl ethidium bromide was visualized under UV illumination to verify RNA loading in each lane; RNA was isolated from total cellular extract (total) and ribosome-enriched subcellular fraction (ribo) prepared from RoSH2 cells before and 48 hours after induction of endothelial differentiation by plating matrigel-coated plates; b) Distribution of rpL5 mRNA in total and ribosome-enriched RNAs before and after differentiation. RT-PCR using oligo-dT-primed cDNA, and Rpl5 and Fkbp12 specific primers was performed on 10 fold serial dilution i.e. 1×, 10× and 100× of RNAs.; c) RT-PCR analysis of transcript abundance in RoSH2 cells before and after differentiation. RoSH2 cells were induced to undergo endothelial differentiation by plating on matrigel. Total and ribosome-enriched RNA were purified at 0 and 48 hours and analyzed by RT-PCR for transcript abundance of G1 cyclins and endothelial receptors; d) Western blot analysis. RoSH2 cells were induced to undergo endothelial differentiation by plating on matrigel and at 0, 6, 24 and 48 hours, cell lysates were prepared and assayed by western blot analysis. Tsc2 protein was used as an internal control for loading between lanes and between blots; e) Effect of rapamycin on Rpl5, cyclinD2 and Tek transcript abundance in total and ribosome-enriched RNA. Total and ribosome-enriched RNA were isolated from RoSH2 cells treated for 0 and 48 hours with 50 ηM rapamycin treatment and analyzed by RT-PCR; f) Western blot analysis. Cell lysates from rapamycin-treated RoSH2 cells at 0, 6, 24 and 48 hours were assayed by western blot analysis for cyclinD2 and Tek. Tsc2 protein was used as an internal control for loading between lanes and between blots.

    Journal: Journal of Molecular Signaling

    Article Title: PI3 K/Akt/mTOR-mediated translational control regulates proliferation and differentiation of lineage-restricted RoSH stem cell lines

    doi: 10.1186/1750-2187-2-9

    Figure Lengend Snippet: Translational control during endothelial differentiation and rapamycin treatment . a) Preparation of total RNA and ribosome-enriched RNA. RoSH2 cells, RoSH2 induced to differentiate by plating on matrigel for 48 hours and RoSH2 cells treated with 50 ηM rapamycin for 48 hours were harvested. Total and ribosome-enriched RNAs were quantitated by absorbance at 260 nm. 2 μg of RNA from each of the two cellular fractions were separated on a 1.0% agarose gel, stained with ethidium bromide and visualized under UV illumination. Top panel Representative RNA samples from total and ribosome-enriched RNAs prepared from undifferentiated (Undiff), differentiated (Diff) and rapamycin-treated (Rapa) RoSH2 cells are shown. Bottom panel For each sample, 1 μl of serially diluted RNA sample mixed with 1 μl of 0.5 ηg/μl ethidium bromide was visualized under UV illumination to verify RNA loading in each lane; RNA was isolated from total cellular extract (total) and ribosome-enriched subcellular fraction (ribo) prepared from RoSH2 cells before and 48 hours after induction of endothelial differentiation by plating matrigel-coated plates; b) Distribution of rpL5 mRNA in total and ribosome-enriched RNAs before and after differentiation. RT-PCR using oligo-dT-primed cDNA, and Rpl5 and Fkbp12 specific primers was performed on 10 fold serial dilution i.e. 1×, 10× and 100× of RNAs.; c) RT-PCR analysis of transcript abundance in RoSH2 cells before and after differentiation. RoSH2 cells were induced to undergo endothelial differentiation by plating on matrigel. Total and ribosome-enriched RNA were purified at 0 and 48 hours and analyzed by RT-PCR for transcript abundance of G1 cyclins and endothelial receptors; d) Western blot analysis. RoSH2 cells were induced to undergo endothelial differentiation by plating on matrigel and at 0, 6, 24 and 48 hours, cell lysates were prepared and assayed by western blot analysis. Tsc2 protein was used as an internal control for loading between lanes and between blots; e) Effect of rapamycin on Rpl5, cyclinD2 and Tek transcript abundance in total and ribosome-enriched RNA. Total and ribosome-enriched RNA were isolated from RoSH2 cells treated for 0 and 48 hours with 50 ηM rapamycin treatment and analyzed by RT-PCR; f) Western blot analysis. Cell lysates from rapamycin-treated RoSH2 cells at 0, 6, 24 and 48 hours were assayed by western blot analysis for cyclinD2 and Tek. Tsc2 protein was used as an internal control for loading between lanes and between blots.

    Article Snippet: Primary antibodies were 1:200 dilution of rabbit anti-phosphoRps6kb1, Rps6kb1, Eif4ebp1, Tsc2, phosphoAkt1, cyclinD1, cyclinD2, cyclinD3 and cyclinE1, goat anti-cyclinE2 and anti-(4E-BP1) polyclonal antibodies (Santa Cruz Biotechnology, CA), and 1:500 dilution of anti-MAPK (p42/44) rabbit polyclonal antibody (Cell Signaling Technology, MA), and rat anti-Kdr and rabbit anti-Tek (PharMingen, MA).

    Techniques: Agarose Gel Electrophoresis, Staining, Isolation, Reverse Transcription Polymerase Chain Reaction, Serial Dilution, Purification, Western Blot

    (A-C), in situ hybridisation for miR-133a-3p expression in normal (C) and diseased (A and B) retinas. (D), luciferase assay to study the correlation of miR-133a-3p with cyclin D2 expression in 661 photoreceptor cells. The expression of miR-133a-3p in the outer nuclear layer (ONL) was increased after induced Müller cell ablation. Co-transfection of 661w cells using a vector containing 3’UTR region of the cyclin D2 gene along with a miR-133a-3p mimic (Vector + miR) induced a significant increase in the normalised firefly luciferase activity compared with cells co-transfected using the vector with a control miRNA (Vector + Ctrl) (*P

    Journal: PLoS ONE

    Article Title: Profiling of MicroRNAs Involved in Retinal Degeneration Caused by Selective Müller Cell Ablation

    doi: 10.1371/journal.pone.0118949

    Figure Lengend Snippet: (A-C), in situ hybridisation for miR-133a-3p expression in normal (C) and diseased (A and B) retinas. (D), luciferase assay to study the correlation of miR-133a-3p with cyclin D2 expression in 661 photoreceptor cells. The expression of miR-133a-3p in the outer nuclear layer (ONL) was increased after induced Müller cell ablation. Co-transfection of 661w cells using a vector containing 3’UTR region of the cyclin D2 gene along with a miR-133a-3p mimic (Vector + miR) induced a significant increase in the normalised firefly luciferase activity compared with cells co-transfected using the vector with a control miRNA (Vector + Ctrl) (*P

    Article Snippet: Cleaved-caspase 9, cyclin D2 and JAK2, were mainly expressed in the ONL in areas corresponding to Müller cell ablation, whereas upregulation of calmodulin was observed in the outer plexiform layer, ONL and photoreceptor segments after Müller cell ablation.

    Techniques: In Situ, Hybridization, Expressing, Luciferase, Cotransfection, Plasmid Preparation, Activity Assay, Transfection

    Nox4 overexpression activates c-myc resulting in increased cyclin D2 expression. (A) Representative immunoblot showing increased c-myc phosphorylation in 2 week-old Nox4 Tg hearts compared with Wt littermate controls. The histogram represents the ratio of phosphorylated c-myc to total c-myc protein levels (n = 3). (B) Representative immunoblot and quantitative histogram showing phosphorylated and total c-myc levels in NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence or absence of PD98059 as indicated (n = 3). (C) Q-PCR analyses of c-myc mRNA expression, relative to that of β-actin (n = 3), and (D) representative immunoblot indicating c-myc protein expression, in NRCs after 48 h of c-myc siRNA treatment as indicated. β-Actin is shown as a loading control for protein expression. (E) Representative immunoblot and quantitative histogram showing cyclin D2 protein expression, relative to that of β-actin, in NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence of scrambled or c-myc siRNA as indicated (n = 3). All data are presented as mean ± S.E. * P

    Journal: Journal of Molecular and Cellular Cardiology

    Article Title: Redox regulation of cardiomyocyte cell cycling via an ERK1/2 and c-Myc-dependent activation of cyclin D2 transcription

    doi: 10.1016/j.yjmcc.2014.10.017

    Figure Lengend Snippet: Nox4 overexpression activates c-myc resulting in increased cyclin D2 expression. (A) Representative immunoblot showing increased c-myc phosphorylation in 2 week-old Nox4 Tg hearts compared with Wt littermate controls. The histogram represents the ratio of phosphorylated c-myc to total c-myc protein levels (n = 3). (B) Representative immunoblot and quantitative histogram showing phosphorylated and total c-myc levels in NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence or absence of PD98059 as indicated (n = 3). (C) Q-PCR analyses of c-myc mRNA expression, relative to that of β-actin (n = 3), and (D) representative immunoblot indicating c-myc protein expression, in NRCs after 48 h of c-myc siRNA treatment as indicated. β-Actin is shown as a loading control for protein expression. (E) Representative immunoblot and quantitative histogram showing cyclin D2 protein expression, relative to that of β-actin, in NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence of scrambled or c-myc siRNA as indicated (n = 3). All data are presented as mean ± S.E. * P

    Article Snippet: By contrast, in the Tg mouse hearts, there remain significant levels of cyclin D2 at 11 days after birth ( G, H).

    Techniques: Over Expression, Expressing, Transduction, Polymerase Chain Reaction

    Nox4-generated ROS increases the rate of cyclin D2 transcription via a highly conserved, cis -acting promoter fragment. (A) A DNA sequence comparison showing high conservation between regions of the proximal human and mouse cyclin D2 promoter regions, upstream of exon 1. Functional binding sites for indicated transcription factors [37,48] are indicated and shaded in grey. Stars indicate nucleotide homology between species. Nucleotides are numbered relative to start of translation of each gene. (B) Relative luciferase activity (RLA) resulting from a promoterless firefly luciferase vector (empty vector) or from a construct comprising a 1766 bp genomic fragment of the proximal mouse cyclin D2 promoter ligated upstream of the luciferase reporter gene (cyclin D2-luc) in NRCs transduced with either AdNox4 or AdβGal for 28 h. (C D) RLA resulting from the cyclin D2-luc in NRCs transduced with either AdNox4 or AdβGal for 28 h in the presence of scrambled or cmyc siRNA(C), or PD90859 (D), as indicated. In each case, luciferase activities are normalised to that of a co-transfected Renilla luciferase vector and expressed as relative luminescence units (RLUs; n = 3). The RLU resulting from cyclin D2-luc transduced with AdβGal was set to 1.0 in each case. All data are presented as mean ± S.E. * P

    Journal: Journal of Molecular and Cellular Cardiology

    Article Title: Redox regulation of cardiomyocyte cell cycling via an ERK1/2 and c-Myc-dependent activation of cyclin D2 transcription

    doi: 10.1016/j.yjmcc.2014.10.017

    Figure Lengend Snippet: Nox4-generated ROS increases the rate of cyclin D2 transcription via a highly conserved, cis -acting promoter fragment. (A) A DNA sequence comparison showing high conservation between regions of the proximal human and mouse cyclin D2 promoter regions, upstream of exon 1. Functional binding sites for indicated transcription factors [37,48] are indicated and shaded in grey. Stars indicate nucleotide homology between species. Nucleotides are numbered relative to start of translation of each gene. (B) Relative luciferase activity (RLA) resulting from a promoterless firefly luciferase vector (empty vector) or from a construct comprising a 1766 bp genomic fragment of the proximal mouse cyclin D2 promoter ligated upstream of the luciferase reporter gene (cyclin D2-luc) in NRCs transduced with either AdNox4 or AdβGal for 28 h. (C D) RLA resulting from the cyclin D2-luc in NRCs transduced with either AdNox4 or AdβGal for 28 h in the presence of scrambled or cmyc siRNA(C), or PD90859 (D), as indicated. In each case, luciferase activities are normalised to that of a co-transfected Renilla luciferase vector and expressed as relative luminescence units (RLUs; n = 3). The RLU resulting from cyclin D2-luc transduced with AdβGal was set to 1.0 in each case. All data are presented as mean ± S.E. * P

    Article Snippet: By contrast, in the Tg mouse hearts, there remain significant levels of cyclin D2 at 11 days after birth ( G, H).

    Techniques: Generated, Sequencing, Functional Assay, Binding Assay, Luciferase, Activity Assay, Plasmid Preparation, Construct, Transduction, Transfection

    Nox4-induced proliferation in NRCs is mediated by the transcriptional upregulation of cyclin D2. (A) Q-PCR analyses showing relative cyclin D2 mRNA expression, normalised to β-actin (n = 3) and (B) representative immunoblots and quantitative histograms of protein expression levels normalised to those of laminin A/C in NRCs transduced with either Nox4 (AdNox4) or β-galactosidase (AdβGal) as control, for 30 h (n = 3). (C) Q-PCR analyses of cyclin D2 mRNA levels in NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence or absence of the MEK inhibitor, PD98059 (n = 3). (D) Representative immunoblot and quantitative histograms of cyclin D2 levels (relative to those of β-actin) in protein extracted from cells as described in (C) (n = 3). (E) Representative immunoblot and quantitative histogram of cyclin D2 protein in NRCs after either scrambled (siScr) or cyclin D2 (siD2) siRNA treatment (n = 3). (F) MTS-based cell proliferation assay of NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence of scrambled, or cyclin D2 siRNA (n = 3). All data are presented as mean ± S.E. * P

    Journal: Journal of Molecular and Cellular Cardiology

    Article Title: Redox regulation of cardiomyocyte cell cycling via an ERK1/2 and c-Myc-dependent activation of cyclin D2 transcription

    doi: 10.1016/j.yjmcc.2014.10.017

    Figure Lengend Snippet: Nox4-induced proliferation in NRCs is mediated by the transcriptional upregulation of cyclin D2. (A) Q-PCR analyses showing relative cyclin D2 mRNA expression, normalised to β-actin (n = 3) and (B) representative immunoblots and quantitative histograms of protein expression levels normalised to those of laminin A/C in NRCs transduced with either Nox4 (AdNox4) or β-galactosidase (AdβGal) as control, for 30 h (n = 3). (C) Q-PCR analyses of cyclin D2 mRNA levels in NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence or absence of the MEK inhibitor, PD98059 (n = 3). (D) Representative immunoblot and quantitative histograms of cyclin D2 levels (relative to those of β-actin) in protein extracted from cells as described in (C) (n = 3). (E) Representative immunoblot and quantitative histogram of cyclin D2 protein in NRCs after either scrambled (siScr) or cyclin D2 (siD2) siRNA treatment (n = 3). (F) MTS-based cell proliferation assay of NRCs transduced with either AdNox4 or AdβGal for 30 h in the presence of scrambled, or cyclin D2 siRNA (n = 3). All data are presented as mean ± S.E. * P

    Article Snippet: By contrast, in the Tg mouse hearts, there remain significant levels of cyclin D2 at 11 days after birth ( G, H).

    Techniques: Polymerase Chain Reaction, Expressing, Western Blot, Transduction, Proliferation Assay

    Nox4 overexpression induces the upregulation of cyclin D2 in vivo. (A) Expression levels of cyclin D2 protein during development from Wt mice at embryonic day 14.5, birth (day 0), and 7 days and 14 days postnatal. (B) Q-PCR analysis of the relative expression of endogenous cyclin D2 mRNA in 2 week old Wt and Tg mouse hearts. Triplicate, independent Tg and Wt littermate controls were analysed, and relative expression was normalised to β-actin in all cases. (C) Protein expression levels of cyclin D2 in Wt and Tg hearts at 2 weeks of age. The histogram depicts the mean of triplicate, independent samples normalised to β − actin and expressed in arbitrary units (A.U.). (D) Q-PCR analysis of the relative mRNA expression of the indicated cell cycle-related proteins, normalised to β-actin from 2 week old Wt and Tg mice (n = 3 in each case). (E) Representative immunoblots and quantitative histograms of protein expression levels normalised to β-actin of the indicated cell cycle-related proteins in 2 week old Wt and Tg mice (n = 3 in all cases). All data are presented as mean ± S.E. * P

    Journal: Journal of Molecular and Cellular Cardiology

    Article Title: Redox regulation of cardiomyocyte cell cycling via an ERK1/2 and c-Myc-dependent activation of cyclin D2 transcription

    doi: 10.1016/j.yjmcc.2014.10.017

    Figure Lengend Snippet: Nox4 overexpression induces the upregulation of cyclin D2 in vivo. (A) Expression levels of cyclin D2 protein during development from Wt mice at embryonic day 14.5, birth (day 0), and 7 days and 14 days postnatal. (B) Q-PCR analysis of the relative expression of endogenous cyclin D2 mRNA in 2 week old Wt and Tg mouse hearts. Triplicate, independent Tg and Wt littermate controls were analysed, and relative expression was normalised to β-actin in all cases. (C) Protein expression levels of cyclin D2 in Wt and Tg hearts at 2 weeks of age. The histogram depicts the mean of triplicate, independent samples normalised to β − actin and expressed in arbitrary units (A.U.). (D) Q-PCR analysis of the relative mRNA expression of the indicated cell cycle-related proteins, normalised to β-actin from 2 week old Wt and Tg mice (n = 3 in each case). (E) Representative immunoblots and quantitative histograms of protein expression levels normalised to β-actin of the indicated cell cycle-related proteins in 2 week old Wt and Tg mice (n = 3 in all cases). All data are presented as mean ± S.E. * P

    Article Snippet: By contrast, in the Tg mouse hearts, there remain significant levels of cyclin D2 at 11 days after birth ( G, H).

    Techniques: Over Expression, In Vivo, Expressing, Mouse Assay, Polymerase Chain Reaction, Western Blot

    The expression of D-cyclins in mantle cell lymphoma The differential expression of D-cyclin staining in two case examples of mantle cell lymphoma is shown. Case 1 shows intense cyclin D1 and weak cyclin D2 staining. Case 2 shows weak cyclin D1 and strong cyclin D2 staining. Both cases lack staining for cyclin D3.

    Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

    Article Title: Characterization of D-Cyclin Proteins in Hematolymphoid Neoplasms: Lack of Specificity of Cyclins D2 and D3 Expression in Lymphoma Subtypes

    doi: 10.1038/modpathol.2009.173

    Figure Lengend Snippet: The expression of D-cyclins in mantle cell lymphoma The differential expression of D-cyclin staining in two case examples of mantle cell lymphoma is shown. Case 1 shows intense cyclin D1 and weak cyclin D2 staining. Case 2 shows weak cyclin D1 and strong cyclin D2 staining. Both cases lack staining for cyclin D3.

    Article Snippet: Antibodies directed against D-cyclins were used at a dilution of 1:100 for anti-cyclin D1 (clone SP4, Thermo Fisher Scientific, Fremont, CA), 1:400 for anti-cyclin D2 (clone M-20, Santa Cruz Biotechnology, Santa Cruz, CA), and 1:30 for anti-cyclin D3 (clone DCS-22, Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Expressing, Staining

    Hierarchical cluster analysis of immunohistologic data in diffuse large B-cell lymphoma Hierarchical cluster analysis shows that the expression profiles of cyclin D2 and D3 proteins are similar to each other across the 143 cases of diffuse large B-cell lymphoma, but are distinct from that of germinal center-associated markers CD10, BCL6, HGAL and LMO2, and from non-germinal center markers, BCL2 and MUM1/IRF4.

    Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

    Article Title: Characterization of D-Cyclin Proteins in Hematolymphoid Neoplasms: Lack of Specificity of Cyclins D2 and D3 Expression in Lymphoma Subtypes

    doi: 10.1038/modpathol.2009.173

    Figure Lengend Snippet: Hierarchical cluster analysis of immunohistologic data in diffuse large B-cell lymphoma Hierarchical cluster analysis shows that the expression profiles of cyclin D2 and D3 proteins are similar to each other across the 143 cases of diffuse large B-cell lymphoma, but are distinct from that of germinal center-associated markers CD10, BCL6, HGAL and LMO2, and from non-germinal center markers, BCL2 and MUM1/IRF4.

    Article Snippet: Antibodies directed against D-cyclins were used at a dilution of 1:100 for anti-cyclin D1 (clone SP4, Thermo Fisher Scientific, Fremont, CA), 1:400 for anti-cyclin D2 (clone M-20, Santa Cruz Biotechnology, Santa Cruz, CA), and 1:30 for anti-cyclin D3 (clone DCS-22, Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Expressing

    Cyclin D2 expression in hematopoietic neoplasia Cyclin D2 staining is shown in examples of diffuse large B-cell lymphoma (A), follicular lymphoma (B), marginal zone lymphoma (C), small lymphocytic lymphoma (D), classical Hodgkin lymphoma (E), and peripheral T-cell lymphoma (F). In all cases, the staining is primarily localized to the nucleus, although associated cytoplasmic staining is also seen in some instances.

    Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

    Article Title: Characterization of D-Cyclin Proteins in Hematolymphoid Neoplasms: Lack of Specificity of Cyclins D2 and D3 Expression in Lymphoma Subtypes

    doi: 10.1038/modpathol.2009.173

    Figure Lengend Snippet: Cyclin D2 expression in hematopoietic neoplasia Cyclin D2 staining is shown in examples of diffuse large B-cell lymphoma (A), follicular lymphoma (B), marginal zone lymphoma (C), small lymphocytic lymphoma (D), classical Hodgkin lymphoma (E), and peripheral T-cell lymphoma (F). In all cases, the staining is primarily localized to the nucleus, although associated cytoplasmic staining is also seen in some instances.

    Article Snippet: Antibodies directed against D-cyclins were used at a dilution of 1:100 for anti-cyclin D1 (clone SP4, Thermo Fisher Scientific, Fremont, CA), 1:400 for anti-cyclin D2 (clone M-20, Santa Cruz Biotechnology, Santa Cruz, CA), and 1:30 for anti-cyclin D3 (clone DCS-22, Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Expressing, Staining

    Cyclin D2 expression in normal hematopoietic tissue Cyclin D2 staining is found in a subset of cells in the germinal center, mantle and marginal zones and in the paracortex of a normal tonsil (A and B); double immunohistochemical labeling for cyclin D2 (brown) and CD20 (red) show co-localization of staining in a subset of B-cell (arrows, C); double immunohistochemical labeling for cyclin D2 (red) and CD3 (brown) show co-localization of staining in a subset of T-cell (arrow, D); scattered cells in the cortex and medulla of the normal thymus show cyclin D2-positive cells (E); the normal spleen shows staining in scattered lymphoid cells in the red and white pulp and in endothelial cells but is lacking in splenic marginal zones (F); the normal bone marrow shows staining in scattered lymphoid and plasma cells but is not found in a significant proportion of erythroid or myeloid precursors or megakaryocytes (G).

    Journal: Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc

    Article Title: Characterization of D-Cyclin Proteins in Hematolymphoid Neoplasms: Lack of Specificity of Cyclins D2 and D3 Expression in Lymphoma Subtypes

    doi: 10.1038/modpathol.2009.173

    Figure Lengend Snippet: Cyclin D2 expression in normal hematopoietic tissue Cyclin D2 staining is found in a subset of cells in the germinal center, mantle and marginal zones and in the paracortex of a normal tonsil (A and B); double immunohistochemical labeling for cyclin D2 (brown) and CD20 (red) show co-localization of staining in a subset of B-cell (arrows, C); double immunohistochemical labeling for cyclin D2 (red) and CD3 (brown) show co-localization of staining in a subset of T-cell (arrow, D); scattered cells in the cortex and medulla of the normal thymus show cyclin D2-positive cells (E); the normal spleen shows staining in scattered lymphoid cells in the red and white pulp and in endothelial cells but is lacking in splenic marginal zones (F); the normal bone marrow shows staining in scattered lymphoid and plasma cells but is not found in a significant proportion of erythroid or myeloid precursors or megakaryocytes (G).

    Article Snippet: Antibodies directed against D-cyclins were used at a dilution of 1:100 for anti-cyclin D1 (clone SP4, Thermo Fisher Scientific, Fremont, CA), 1:400 for anti-cyclin D2 (clone M-20, Santa Cruz Biotechnology, Santa Cruz, CA), and 1:30 for anti-cyclin D3 (clone DCS-22, Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Expressing, Staining, Immunohistochemistry, Labeling

    Overexpression of cyclin D2 increases β cell proliferation and β cell mass. (A) Representative immunofluorescence staining for insulin (green), Ki67 (red), and DAPI (blue). White arrows indicate Ins + Ki67 + cells. (B) Quantification of the percentage of β cells expressing Ki67 in WT and KI mice at 3 weeks and 3 months of age. (C) Quantification of β cell mass in WT and KI mice at 3 weeks and 3 months of age. Data shown as mean ± SD (n = 3–4 mice per group). *P

    Journal: Cell Cycle

    Article Title: Cyclin D2 is sufficient to drive β cell self-renewal and regeneration

    doi: 10.1080/15384101.2017.1319999

    Figure Lengend Snippet: Overexpression of cyclin D2 increases β cell proliferation and β cell mass. (A) Representative immunofluorescence staining for insulin (green), Ki67 (red), and DAPI (blue). White arrows indicate Ins + Ki67 + cells. (B) Quantification of the percentage of β cells expressing Ki67 in WT and KI mice at 3 weeks and 3 months of age. (C) Quantification of β cell mass in WT and KI mice at 3 weeks and 3 months of age. Data shown as mean ± SD (n = 3–4 mice per group). *P

    Article Snippet: The membranes were probed with specific antibodies against cyclin D2 (Santa Cruz), and β-tubulin (Sigma-Aldrich).

    Techniques: Over Expression, Immunofluorescence, Staining, Expressing, Mouse Assay

    β cell specific overexpression of cyclin D2 increases cyclin D2 protein levels. (A) Schematic of the alleles used to create RIP-Cre;cycD2;ROSA26 mT/mG mice (KI mice). Black triangles indicate loxP sites. (B) Representative immunofluorescence staining for insulin or dTomato (red), GFP (green), and DAPI (blue) showing efficient Cre recombinase-mediated recombination in KI β cells. (C) Representative immunofluorescence staining for of cyclin D2 (red) and insulin (green) in WT and KI pancreatic sections. (D) Western blot (left panel) and densitometric quantification of cyclin D2 levels (right panel) in isolated islets from WT and KI mice. Data shown as mean ± SD of 3 independent experiments. ** P

    Journal: Cell Cycle

    Article Title: Cyclin D2 is sufficient to drive β cell self-renewal and regeneration

    doi: 10.1080/15384101.2017.1319999

    Figure Lengend Snippet: β cell specific overexpression of cyclin D2 increases cyclin D2 protein levels. (A) Schematic of the alleles used to create RIP-Cre;cycD2;ROSA26 mT/mG mice (KI mice). Black triangles indicate loxP sites. (B) Representative immunofluorescence staining for insulin or dTomato (red), GFP (green), and DAPI (blue) showing efficient Cre recombinase-mediated recombination in KI β cells. (C) Representative immunofluorescence staining for of cyclin D2 (red) and insulin (green) in WT and KI pancreatic sections. (D) Western blot (left panel) and densitometric quantification of cyclin D2 levels (right panel) in isolated islets from WT and KI mice. Data shown as mean ± SD of 3 independent experiments. ** P

    Article Snippet: The membranes were probed with specific antibodies against cyclin D2 (Santa Cruz), and β-tubulin (Sigma-Aldrich).

    Techniques: Over Expression, Mouse Assay, Immunofluorescence, Staining, Western Blot, Isolation

    Overexpression of cyclin D2 enhances β cell replication in old KI mice. (A, B) Representative immunofluorescence staining (A) and quantification (B) for insulin (green), Ki67 (red), and DAPI (blue) in young (6-week-old) WT and KI mice challenged with a single dose of STZ (90 mg/kg). C, D. Representative immunofluorescence staining (C) and quantification (D) for insulin (green), Ki67 (red), and DAPI (blue) in old (8-month-old) WT and KI mice challenged with a single dose of STZ (90 mg/kg). White arrows indicate Ins + Ki67 + cells. Data shown as mean ± SD (n = 3–4 mice per group). *P

    Journal: Cell Cycle

    Article Title: Cyclin D2 is sufficient to drive β cell self-renewal and regeneration

    doi: 10.1080/15384101.2017.1319999

    Figure Lengend Snippet: Overexpression of cyclin D2 enhances β cell replication in old KI mice. (A, B) Representative immunofluorescence staining (A) and quantification (B) for insulin (green), Ki67 (red), and DAPI (blue) in young (6-week-old) WT and KI mice challenged with a single dose of STZ (90 mg/kg). C, D. Representative immunofluorescence staining (C) and quantification (D) for insulin (green), Ki67 (red), and DAPI (blue) in old (8-month-old) WT and KI mice challenged with a single dose of STZ (90 mg/kg). White arrows indicate Ins + Ki67 + cells. Data shown as mean ± SD (n = 3–4 mice per group). *P

    Article Snippet: The membranes were probed with specific antibodies against cyclin D2 (Santa Cruz), and β-tubulin (Sigma-Aldrich).

    Techniques: Over Expression, Mouse Assay, Immunofluorescence, Staining

    Overexpression of cyclin D2 in β cells does not diminish β cell function. (A) Glucose tolerance test was performed in 3-month-old WT and KI mice. (B) Insulin tolerance was performed in 3-month-old WT and KI mice. (C) Glucose stimulated insulin secretion was measured in 3-month-old WT and KI mice. n = 4–5 animals per group. Data shown as mean ± SD of 3 independent experiments. **P

    Journal: Cell Cycle

    Article Title: Cyclin D2 is sufficient to drive β cell self-renewal and regeneration

    doi: 10.1080/15384101.2017.1319999

    Figure Lengend Snippet: Overexpression of cyclin D2 in β cells does not diminish β cell function. (A) Glucose tolerance test was performed in 3-month-old WT and KI mice. (B) Insulin tolerance was performed in 3-month-old WT and KI mice. (C) Glucose stimulated insulin secretion was measured in 3-month-old WT and KI mice. n = 4–5 animals per group. Data shown as mean ± SD of 3 independent experiments. **P

    Article Snippet: The membranes were probed with specific antibodies against cyclin D2 (Santa Cruz), and β-tubulin (Sigma-Aldrich).

    Techniques: Over Expression, Cell Function Assay, Mouse Assay

    Reconstituting cyclin D2 expression restores β cell mass, function, and regenerative capacity. (A) Representative immunofluorescence staining glucagon (red), insulin (green) and DAPI (blue) in 6-week-old WT, KO, and KIKO mice. (B) Quantification of β cell mass in 6-week-old WT, KO, and KIKO mice. (C) GTT was performed in 3-month-old WT, KO, and KIKO mice. (D, E) Representative immunofluorescence staining (F) and quantification (E) for insulin (green), Ki67 (red), and DAPI (blue) in 6-week-old WT, KO and KIKO mice treated with a single dose of STZ (90 mg/kg). White arrows indicate Ins + Ki67 + cells. Data shown as mean ± SD (n = 3 mice per group). * P

    Journal: Cell Cycle

    Article Title: Cyclin D2 is sufficient to drive β cell self-renewal and regeneration

    doi: 10.1080/15384101.2017.1319999

    Figure Lengend Snippet: Reconstituting cyclin D2 expression restores β cell mass, function, and regenerative capacity. (A) Representative immunofluorescence staining glucagon (red), insulin (green) and DAPI (blue) in 6-week-old WT, KO, and KIKO mice. (B) Quantification of β cell mass in 6-week-old WT, KO, and KIKO mice. (C) GTT was performed in 3-month-old WT, KO, and KIKO mice. (D, E) Representative immunofluorescence staining (F) and quantification (E) for insulin (green), Ki67 (red), and DAPI (blue) in 6-week-old WT, KO and KIKO mice treated with a single dose of STZ (90 mg/kg). White arrows indicate Ins + Ki67 + cells. Data shown as mean ± SD (n = 3 mice per group). * P

    Article Snippet: The membranes were probed with specific antibodies against cyclin D2 (Santa Cruz), and β-tubulin (Sigma-Aldrich).

    Techniques: Expressing, Immunofluorescence, Staining, Mouse Assay

    Cyclin D2 is enriched in the ventral peripheral retina during the temporal window of ipsilateral RGC genesis. IHC in coronal cryosections of E11.5 ( A ), E12.5 ( B ), E14.5 ( C ), E15.5 ( D ), and E16.5 ( E ) retina. Cyclin D2 is expressed in the lens and the retinal marginal zone bordering RGCs (labeled with Brn3). Cyclin D2 is highly expressed in the ventral periphery compared with the dorsal periphery within neural retina (white arrows). The cyclin D2 and Brn3 domains are separated by a large gap at E11.5 ( A ). At E13.5 ( B ) and E14.5 ( C ), cyclin D2 + cells intermingle with Brn3 + RGCs only in ventral retina (yellow arrows point to boundary) and have sharp boundaries in dorsal retina. At E16.5 ( E ), cyclin D2 expression is reduced in the retinal periphery and is no longer asymmetric in dorsal and ventral retinal (white arrows). Scale bars, 100 μm.

    Journal: eNeuro

    Article Title: Ipsilateral and Contralateral Retinal Ganglion Cells Express Distinct Genes during Decussation at the Optic Chiasm

    doi: 10.1523/ENEURO.0169-16.2016

    Figure Lengend Snippet: Cyclin D2 is enriched in the ventral peripheral retina during the temporal window of ipsilateral RGC genesis. IHC in coronal cryosections of E11.5 ( A ), E12.5 ( B ), E14.5 ( C ), E15.5 ( D ), and E16.5 ( E ) retina. Cyclin D2 is expressed in the lens and the retinal marginal zone bordering RGCs (labeled with Brn3). Cyclin D2 is highly expressed in the ventral periphery compared with the dorsal periphery within neural retina (white arrows). The cyclin D2 and Brn3 domains are separated by a large gap at E11.5 ( A ). At E13.5 ( B ) and E14.5 ( C ), cyclin D2 + cells intermingle with Brn3 + RGCs only in ventral retina (yellow arrows point to boundary) and have sharp boundaries in dorsal retina. At E16.5 ( E ), cyclin D2 expression is reduced in the retinal periphery and is no longer asymmetric in dorsal and ventral retinal (white arrows). Scale bars, 100 μm.

    Article Snippet: Antibodies The following primary antibodies were used: rabbit anti-Zic2 (RRID:AB_2315623, gift of Stephen Brown, 1:10,000), mouse anti-Islet1/2 (cat. # 39.4D5, RRID:AB_528173, gift of Thomas Jessell, 1:50; this antibody was raised against Islet1 but also reacts with Islet2), rabbit anti-cyclin D2 (Santa Cruz Biotechnology, Santa Cruz, CA, cat. # sc-452, RRID:AB_627350, 1:1000), rat anti-cyclin D2 (Santa Cruz Biotechnology, cat. # sc-593 RRID:AB_2070794, 1:50), goat anti-Brn3 (Santa Cruz Biotechnology, cat. # sc-6026 RRID:AB_673441, 1:200), and mouse anti-Brn3a (EMD Millipore, Billerica, MA, cat. # MAB1585 RRID:AB_94166, 1:200).

    Techniques: Immunohistochemistry, Labeling, Expressing

    Sox2, Math5, and cyclin D2 are enriched in the ipsilateral RGC population in addition to progenitor cells throughout the retina. ISH analysis at E15.5. A , Sox2 mRNA is expressed in the neuroblastic layer and Zic2 + RGCs but not Zic2 – /Islet1/2 + (differentiated) RGCs extra-VT retina. B , Similarly, Math5 mRNA is expressed in Zic2 + RGCs located at the periphery of the VT retina in the RGC layer as well as in RGC precursors in the neuroblastic layer (red arrows). C , Cyclin D2 mRNA is expressed in the basal process of cells within the peripheral margin of the retina, particularly in the ventral retina, and also at low levels in the Zic2 + RGC zone at E15.5. D , The asymmetric expression of cyclin D2, with higher levels in ventral retina, is more pronounced at E13.5. Red brackets mark the Zic2 + ipsilateral RGC domain. Scale bars, 250 μm.

    Journal: eNeuro

    Article Title: Ipsilateral and Contralateral Retinal Ganglion Cells Express Distinct Genes during Decussation at the Optic Chiasm

    doi: 10.1523/ENEURO.0169-16.2016

    Figure Lengend Snippet: Sox2, Math5, and cyclin D2 are enriched in the ipsilateral RGC population in addition to progenitor cells throughout the retina. ISH analysis at E15.5. A , Sox2 mRNA is expressed in the neuroblastic layer and Zic2 + RGCs but not Zic2 – /Islet1/2 + (differentiated) RGCs extra-VT retina. B , Similarly, Math5 mRNA is expressed in Zic2 + RGCs located at the periphery of the VT retina in the RGC layer as well as in RGC precursors in the neuroblastic layer (red arrows). C , Cyclin D2 mRNA is expressed in the basal process of cells within the peripheral margin of the retina, particularly in the ventral retina, and also at low levels in the Zic2 + RGC zone at E15.5. D , The asymmetric expression of cyclin D2, with higher levels in ventral retina, is more pronounced at E13.5. Red brackets mark the Zic2 + ipsilateral RGC domain. Scale bars, 250 μm.

    Article Snippet: Antibodies The following primary antibodies were used: rabbit anti-Zic2 (RRID:AB_2315623, gift of Stephen Brown, 1:10,000), mouse anti-Islet1/2 (cat. # 39.4D5, RRID:AB_528173, gift of Thomas Jessell, 1:50; this antibody was raised against Islet1 but also reacts with Islet2), rabbit anti-cyclin D2 (Santa Cruz Biotechnology, Santa Cruz, CA, cat. # sc-452, RRID:AB_627350, 1:1000), rat anti-cyclin D2 (Santa Cruz Biotechnology, cat. # sc-593 RRID:AB_2070794, 1:50), goat anti-Brn3 (Santa Cruz Biotechnology, cat. # sc-6026 RRID:AB_673441, 1:200), and mouse anti-Brn3a (EMD Millipore, Billerica, MA, cat. # MAB1585 RRID:AB_94166, 1:200).

    Techniques: In Situ Hybridization, Expressing

    Effect of AMPK activation on cyclin D2 mRNA expression in granulosa cells. Granulosa cells from 3-d estradiol primed immature rats were cultured in serum-free, phenol red-free medium overnight followed by incubation with AMPK activator (AICAR) for 1 h.

    Journal: Endocrinology

    Article Title: Follicle-Stimulating Hormone Inhibits Adenosine 5?-Monophosphate-Activated Protein Kinase Activation and Promotes Cell Proliferation of Primary Granulosa Cells in Culture through an Akt-Dependent Pathway

    doi: 10.1210/en.2008-1032

    Figure Lengend Snippet: Effect of AMPK activation on cyclin D2 mRNA expression in granulosa cells. Granulosa cells from 3-d estradiol primed immature rats were cultured in serum-free, phenol red-free medium overnight followed by incubation with AMPK activator (AICAR) for 1 h.

    Article Snippet: The real-time PCR quantification was then performed using 5 μl of the diluted cDNAs in triplicates and predesigned primers and probes for cyclin D2 (TaqMan Assay on Demand gene expression product Applied Biosystems, Foster City, CA).

    Techniques: Activation Assay, Expressing, Cell Culture, Incubation

    Cyclin D/CDK4 negatively regulates PD-L1 protein stability a, b, Immunoblot (IB) analysis of whole cell lysates (WCL) derived from wild type (WT), cyclin A1 −/− A2 −/− or WT, cyclin E1 −/− E2 −/− MEFs. c, Quantitative real-time PCR (qRT-PCR) analysis of relative mRNA levels of PD-L1 from wild type MEFs and cyclin D1 −/− D2 −/− D3 −/− MEFs. Data were represented as mean ± s.d, n = 5. d, Cell cycle profiles for WT and cyclin D1 −/− D2 −/− D3 −/− MEFs, which were labeled with BrdU and analyzed by FACS. e, IB analysis of WCL derived from cyclin D1 fl/fl D2 −/− D3 fl/fl MEFs with or without depleting cyclin D1 and cyclin D3 by pLenti-Cre via viral infection (pLenti-EGFP as a negative control), selected with puromycin (1 μg/ml) for 72 hours before harvesting. f, IB analysis of WCL derived from cyclin D1 −/− D2 −/− D3 −/− MEFs stably reintroducing cyclin D1 , cyclin D2 , or cyclin D3 , respectively, with empty vector (EV) as a negative control. g, IB analysis of WCL derived from mouse mammary tumors induced by MMTV- c-Myc with/without genetic depletion of cyclin D1 . n = 5 mice per experimental group. h, IB analysis of WCL derived from WCL derived from wild type and cdk6 −/− MEFs. i, j, IB analysis of WCL derived from MDA-MB-231 cells stably expressing shCDK6 or shCDK2 as well as shScr as a negative control, respectively. k, l, IB analysis of WCL derived from MDA-MB-231 cells transfected with indicated constructs (k) and the intensity of PD-L1 band was quantified by the ImageJ software (l) . m, IB analysis of WCL derived from MDA-MB-231 cells depleted of Rb (with shScr as a negative control) treated with the CDK4/6 inhibitor, palbociclib, where indicated. n, o, IB analysis of WCL derived from mouse CT26 or 4T1 tumor cell lines treated with or without the CDK4/6 inhibitor, palbociclib or ribociclib, respectively. p, q, IB analysis of WCL derived from MDA-MB-231 cells pre-treated with palbociclib (1 μM) for 36 hours before treatment with cycloheximide (CHX) for the indicated time points (p) and PD-L1 protein abundance was quantified by the ImageJ and plotted as indicated (q) . r, IB analysis of WCL derived from 19 different cancer cell lines with indicated antibodies. s–u, IB analysis of WCL derived from MCF7, T47D or HLF stably expressing p16 as well as EV as a negative control. v–x, IB analysis of WCL derived from MDA-MB-436, BT549 or HCC1937 stably expressing three independent shRNAs against p16 as well as shScr as a negative control.

    Journal: Nature

    Article Title: Cyclin D-CDK4 kinase destabilizes PD-L1 via Cul3SPOP to control cancer immune surveillance

    doi: 10.1038/nature25015

    Figure Lengend Snippet: Cyclin D/CDK4 negatively regulates PD-L1 protein stability a, b, Immunoblot (IB) analysis of whole cell lysates (WCL) derived from wild type (WT), cyclin A1 −/− A2 −/− or WT, cyclin E1 −/− E2 −/− MEFs. c, Quantitative real-time PCR (qRT-PCR) analysis of relative mRNA levels of PD-L1 from wild type MEFs and cyclin D1 −/− D2 −/− D3 −/− MEFs. Data were represented as mean ± s.d, n = 5. d, Cell cycle profiles for WT and cyclin D1 −/− D2 −/− D3 −/− MEFs, which were labeled with BrdU and analyzed by FACS. e, IB analysis of WCL derived from cyclin D1 fl/fl D2 −/− D3 fl/fl MEFs with or without depleting cyclin D1 and cyclin D3 by pLenti-Cre via viral infection (pLenti-EGFP as a negative control), selected with puromycin (1 μg/ml) for 72 hours before harvesting. f, IB analysis of WCL derived from cyclin D1 −/− D2 −/− D3 −/− MEFs stably reintroducing cyclin D1 , cyclin D2 , or cyclin D3 , respectively, with empty vector (EV) as a negative control. g, IB analysis of WCL derived from mouse mammary tumors induced by MMTV- c-Myc with/without genetic depletion of cyclin D1 . n = 5 mice per experimental group. h, IB analysis of WCL derived from WCL derived from wild type and cdk6 −/− MEFs. i, j, IB analysis of WCL derived from MDA-MB-231 cells stably expressing shCDK6 or shCDK2 as well as shScr as a negative control, respectively. k, l, IB analysis of WCL derived from MDA-MB-231 cells transfected with indicated constructs (k) and the intensity of PD-L1 band was quantified by the ImageJ software (l) . m, IB analysis of WCL derived from MDA-MB-231 cells depleted of Rb (with shScr as a negative control) treated with the CDK4/6 inhibitor, palbociclib, where indicated. n, o, IB analysis of WCL derived from mouse CT26 or 4T1 tumor cell lines treated with or without the CDK4/6 inhibitor, palbociclib or ribociclib, respectively. p, q, IB analysis of WCL derived from MDA-MB-231 cells pre-treated with palbociclib (1 μM) for 36 hours before treatment with cycloheximide (CHX) for the indicated time points (p) and PD-L1 protein abundance was quantified by the ImageJ and plotted as indicated (q) . r, IB analysis of WCL derived from 19 different cancer cell lines with indicated antibodies. s–u, IB analysis of WCL derived from MCF7, T47D or HLF stably expressing p16 as well as EV as a negative control. v–x, IB analysis of WCL derived from MDA-MB-436, BT549 or HCC1937 stably expressing three independent shRNAs against p16 as well as shScr as a negative control.

    Article Snippet: Antibodies Anti-PD-L1 (E1L3N) rabbit mAb (13684), anti-pS10-H3 (3377), anti-pS780-Rb (8180), anti-pS807/811-Rb (8516), anti-Rb (9309), anti-cyclin D1 (2978), anti-cyclin D2 (3741), anti-CDK6 (3136), anti-cullin 3 (2759), anti-GST (2625), rabbit polyclonal anti-Myc-Tag antibody (2278) and mouse monoclonal anti-Myc-Tag (2276) antibodies were purchased from Cell Signaling Technology.

    Techniques: Derivative Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Labeling, FACS, Infection, Negative Control, Stable Transfection, Plasmid Preparation, Mouse Assay, Multiple Displacement Amplification, Expressing, Transfection, Construct, Software

    Cyclin D/CDK4-mediated phosphorylation of SPOP at the Ser6 residue promotes its binding with 14-3-3γ to reduce its poly-ubiquitination and subsequent degradation by APC/Cdh1 a, A sequence comparison of conserved SP sites and putative 14-3-3γ binding motif in SPOP. b, Immunoblot (IB) analysis of whole cell lysates (WCL) and immunoprecipitation (IP) derived from 293T cells transfected with indicated constructs and treated with MG132 (10 μM) for 12 hours before harvesting. c, d, In vitro kinase assays with recombinant Rb and SPOP as substrates and cyclin D1/CDK4, cyclin D2/CDK4 and cyclin D3/CDK4 as kinase complex were performed. BSA was used as a negative control where indicated. e, IB analysis of WCL and immunoprecipitation (IP) derived from MDA-MB-231 cells transfected with indicated constructs, which were treated with/without palbociclib (1 μM) for 12 hours. f, Streptavidin beads pull-down assay for biotin-labeled SPOP peptide with/without phosphorylation at the Ser6 residue to examine its in vitro association with 14-3-3γ. g, IB analysis of WCL and GST pull-down precipitates derived from 293T cells transfected with indicated constructs and treated with MG132 (10 μM) for 12 hours before harvesting. h, i, IB analysis of WCL and IP derived from 293T cells transfected with indicated constructs and treated with MG132 (10 μM) for 12 hours before harvesting. j, k, IB analysis of WCL derived from 293T cells transfected with indicated constructs. 36 h post transfection, cells were treated with 20 μg/ml cycloheximide (CHX) as indicated time points (j) . The protein abundance of SPOP-WT and S6A mutant were quantified by the ImageJ software and plotted accordingly (k) . l, p, IB of WCL and Ni-NTA pull-down products derived from the lysates of PC3 cells transfected with the indicated constructs. Cells were treated with MG132 (30 μM) for 6 hours before harvesting and lysed in the denaturing buffer for following assay. m–o, IB analysis of WCL and IP derived from 293T cells transfected with indicated constructs and treated with MG132 (10 μM) and with/without palbociclib (1 μM) for 12 hours before harvesting. q–s, IB of WCLs derived from PC3, BT549 and HeLa cells stably expressing sh14-3-3γ as well as shScr as a negative control. t, IB of WCL derived from HeLa cells stably expressing shScr or sh 14-3-3γ synchronized in M phase by nocodazole treatment prior to releasing back into the cell cycle for the indicated times.

    Journal: Nature

    Article Title: Cyclin D-CDK4 kinase destabilizes PD-L1 via Cul3SPOP to control cancer immune surveillance

    doi: 10.1038/nature25015

    Figure Lengend Snippet: Cyclin D/CDK4-mediated phosphorylation of SPOP at the Ser6 residue promotes its binding with 14-3-3γ to reduce its poly-ubiquitination and subsequent degradation by APC/Cdh1 a, A sequence comparison of conserved SP sites and putative 14-3-3γ binding motif in SPOP. b, Immunoblot (IB) analysis of whole cell lysates (WCL) and immunoprecipitation (IP) derived from 293T cells transfected with indicated constructs and treated with MG132 (10 μM) for 12 hours before harvesting. c, d, In vitro kinase assays with recombinant Rb and SPOP as substrates and cyclin D1/CDK4, cyclin D2/CDK4 and cyclin D3/CDK4 as kinase complex were performed. BSA was used as a negative control where indicated. e, IB analysis of WCL and immunoprecipitation (IP) derived from MDA-MB-231 cells transfected with indicated constructs, which were treated with/without palbociclib (1 μM) for 12 hours. f, Streptavidin beads pull-down assay for biotin-labeled SPOP peptide with/without phosphorylation at the Ser6 residue to examine its in vitro association with 14-3-3γ. g, IB analysis of WCL and GST pull-down precipitates derived from 293T cells transfected with indicated constructs and treated with MG132 (10 μM) for 12 hours before harvesting. h, i, IB analysis of WCL and IP derived from 293T cells transfected with indicated constructs and treated with MG132 (10 μM) for 12 hours before harvesting. j, k, IB analysis of WCL derived from 293T cells transfected with indicated constructs. 36 h post transfection, cells were treated with 20 μg/ml cycloheximide (CHX) as indicated time points (j) . The protein abundance of SPOP-WT and S6A mutant were quantified by the ImageJ software and plotted accordingly (k) . l, p, IB of WCL and Ni-NTA pull-down products derived from the lysates of PC3 cells transfected with the indicated constructs. Cells were treated with MG132 (30 μM) for 6 hours before harvesting and lysed in the denaturing buffer for following assay. m–o, IB analysis of WCL and IP derived from 293T cells transfected with indicated constructs and treated with MG132 (10 μM) and with/without palbociclib (1 μM) for 12 hours before harvesting. q–s, IB of WCLs derived from PC3, BT549 and HeLa cells stably expressing sh14-3-3γ as well as shScr as a negative control. t, IB of WCL derived from HeLa cells stably expressing shScr or sh 14-3-3γ synchronized in M phase by nocodazole treatment prior to releasing back into the cell cycle for the indicated times.

    Article Snippet: Antibodies Anti-PD-L1 (E1L3N) rabbit mAb (13684), anti-pS10-H3 (3377), anti-pS780-Rb (8180), anti-pS807/811-Rb (8516), anti-Rb (9309), anti-cyclin D1 (2978), anti-cyclin D2 (3741), anti-CDK6 (3136), anti-cullin 3 (2759), anti-GST (2625), rabbit polyclonal anti-Myc-Tag antibody (2278) and mouse monoclonal anti-Myc-Tag (2276) antibodies were purchased from Cell Signaling Technology.

    Techniques: Binding Assay, Sequencing, Immunoprecipitation, Derivative Assay, Transfection, Construct, In Vitro, Recombinant, Negative Control, Multiple Displacement Amplification, Pull Down Assay, Labeling, Mutagenesis, Software, Stable Transfection, Expressing

    Inhibition of proliferation of and relevant gene expression in mouse granulosa cells (mGC) by miR-181a. mGC was infected with Ad-miR-181a (multiplicity of infection, MOI = 0, 25, and 50) for 48 h. (A) MiR-181a level was measured by qRT-PCR. (B) Result of a CCK-8 assay examining the proliferation of mGC. (C) qRT-PCR and (D) Western blot analysis of cyclin D2 mRNA and protein levels, respectively, in mGC. (E) Protein level of proliferating cell nuclear antigen (PCNA) as determined by Western blotting. Relative protein levels were measured by densitometry using Quantity One Software and normalized to β-actin, Ad-LacZ group; the ratios were presented above the Western blot bands. *p

    Journal: PLoS ONE

    Article Title: MicroRNA-181a Suppresses Mouse Granulosa Cell Proliferation by Targeting Activin Receptor IIA

    doi: 10.1371/journal.pone.0059667

    Figure Lengend Snippet: Inhibition of proliferation of and relevant gene expression in mouse granulosa cells (mGC) by miR-181a. mGC was infected with Ad-miR-181a (multiplicity of infection, MOI = 0, 25, and 50) for 48 h. (A) MiR-181a level was measured by qRT-PCR. (B) Result of a CCK-8 assay examining the proliferation of mGC. (C) qRT-PCR and (D) Western blot analysis of cyclin D2 mRNA and protein levels, respectively, in mGC. (E) Protein level of proliferating cell nuclear antigen (PCNA) as determined by Western blotting. Relative protein levels were measured by densitometry using Quantity One Software and normalized to β-actin, Ad-LacZ group; the ratios were presented above the Western blot bands. *p

    Article Snippet: Immunoblotting was performed with primary antibodies against cyclin D2 (1∶1,000; Cell Signaling Technology, Danvers, MA, USA), phospho-Smad2 (Ser465/467; 1∶1,000; Cell Signaling Technology; also detects phosphorylated Smad3 at Ser423/425), Smad2 (1∶1,000; Cell Signaling Technology), PCNA (1∶500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), or acvr2a (1∶500; Santa Cruz Biotechnology). β-actin (1∶10,000; Abcam, Cambridge, MA, USA) was measured as an internal control.

    Techniques: Inhibition, Expressing, Infection, Quantitative RT-PCR, CCK-8 Assay, Western Blot, Software

    Effect of miR-181a inhibitor on mouse granulosa cell (mGC) proliferation. mGC was transfected with indicated miR-181a inhibitor (anti-sense oligonucleotide of miR-181a) or miRNA inhibitor negative control (miRNA inhibitor control) for 48 h. (A) MiR-181a expression was measured by qRT-PCR. (B) The proliferation of mGC was examined by CCK-8 after transfection of miR-181a inhibitor. Cyclin D2 mRNA (C) and protein (D) levels measured by qRT-PCR and Western blotting. (E) qRT-PCR and (F) Western blot analysis showed acvr2a mRNA and protein levels in mGC treated with miR-181a inhibitor. Relative protein levels were measured by densitometry using Quantity One Software and normalized to β-actin, the control group; the ratios were presented above the Western blot bands. *p

    Journal: PLoS ONE

    Article Title: MicroRNA-181a Suppresses Mouse Granulosa Cell Proliferation by Targeting Activin Receptor IIA

    doi: 10.1371/journal.pone.0059667

    Figure Lengend Snippet: Effect of miR-181a inhibitor on mouse granulosa cell (mGC) proliferation. mGC was transfected with indicated miR-181a inhibitor (anti-sense oligonucleotide of miR-181a) or miRNA inhibitor negative control (miRNA inhibitor control) for 48 h. (A) MiR-181a expression was measured by qRT-PCR. (B) The proliferation of mGC was examined by CCK-8 after transfection of miR-181a inhibitor. Cyclin D2 mRNA (C) and protein (D) levels measured by qRT-PCR and Western blotting. (E) qRT-PCR and (F) Western blot analysis showed acvr2a mRNA and protein levels in mGC treated with miR-181a inhibitor. Relative protein levels were measured by densitometry using Quantity One Software and normalized to β-actin, the control group; the ratios were presented above the Western blot bands. *p

    Article Snippet: Immunoblotting was performed with primary antibodies against cyclin D2 (1∶1,000; Cell Signaling Technology, Danvers, MA, USA), phospho-Smad2 (Ser465/467; 1∶1,000; Cell Signaling Technology; also detects phosphorylated Smad3 at Ser423/425), Smad2 (1∶1,000; Cell Signaling Technology), PCNA (1∶500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), or acvr2a (1∶500; Santa Cruz Biotechnology). β-actin (1∶10,000; Abcam, Cambridge, MA, USA) was measured as an internal control.

    Techniques: Transfection, Negative Control, Expressing, Quantitative RT-PCR, CCK-8 Assay, Western Blot, Software

    Effect of activin A on miR-181a expression and mouse granulosa cell (mGC) proliferation. mGC was isolated from 21-day-old mouse ovaries. (A) mGC was treated with indicated concentrations of activin A for 24 h. MiR-181a level was determined by qRT-PCR. (B) qRT-PCR analysis was performed to measure miR-181a level in mGC treated with activin A (50 ng/ml) for up to 48 h. (C) The proliferation of mGC was measured by CCK-8 after treated with activin A for 48 h. Cyclin D2 mRNA (D) and protein (E) levels were examined in mGC after treated with activin A (50 ng/ml) for 48 h by qRT-PCR and Western blotting, respectively. Relative protein levels were measured by densitometry using Quantity One Software and normalized to β-actin, the control group; the ratios were presented above the Western blot bands. All experiments were performed three times. *p

    Journal: PLoS ONE

    Article Title: MicroRNA-181a Suppresses Mouse Granulosa Cell Proliferation by Targeting Activin Receptor IIA

    doi: 10.1371/journal.pone.0059667

    Figure Lengend Snippet: Effect of activin A on miR-181a expression and mouse granulosa cell (mGC) proliferation. mGC was isolated from 21-day-old mouse ovaries. (A) mGC was treated with indicated concentrations of activin A for 24 h. MiR-181a level was determined by qRT-PCR. (B) qRT-PCR analysis was performed to measure miR-181a level in mGC treated with activin A (50 ng/ml) for up to 48 h. (C) The proliferation of mGC was measured by CCK-8 after treated with activin A for 48 h. Cyclin D2 mRNA (D) and protein (E) levels were examined in mGC after treated with activin A (50 ng/ml) for 48 h by qRT-PCR and Western blotting, respectively. Relative protein levels were measured by densitometry using Quantity One Software and normalized to β-actin, the control group; the ratios were presented above the Western blot bands. All experiments were performed three times. *p

    Article Snippet: Immunoblotting was performed with primary antibodies against cyclin D2 (1∶1,000; Cell Signaling Technology, Danvers, MA, USA), phospho-Smad2 (Ser465/467; 1∶1,000; Cell Signaling Technology; also detects phosphorylated Smad3 at Ser423/425), Smad2 (1∶1,000; Cell Signaling Technology), PCNA (1∶500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), or acvr2a (1∶500; Santa Cruz Biotechnology). β-actin (1∶10,000; Abcam, Cambridge, MA, USA) was measured as an internal control.

    Techniques: Expressing, Isolation, Quantitative RT-PCR, CCK-8 Assay, Western Blot, Software

    Inactivation of the activin signaling pathway by miR-181a. Western blot analysis of the levels of Smad2 and phosphorylated Smad2 (Ser465/467) in mouse granulosa cells (mGC) treated with 50 ng/ml activin A (A) or infected with 50 MOI Ad-miR-181a (B) for 24 h. (C) mGC was infected with 50 MOI Ad-miR-181a for 24 h, and cells were then treated with 50 ng/ml activin A for another 24 h. The protein level of Smad2 and phosphorylated Smad2 were measured by Western blotting. (D) qRT-PCR analysis of Smad2 and cyclin D2 expression in mGC transfected with 50 nM siRNA duplexes targeting mouse Smad2 (siSmad2) or siRNA negative control for 48 h. *p

    Journal: PLoS ONE

    Article Title: MicroRNA-181a Suppresses Mouse Granulosa Cell Proliferation by Targeting Activin Receptor IIA

    doi: 10.1371/journal.pone.0059667

    Figure Lengend Snippet: Inactivation of the activin signaling pathway by miR-181a. Western blot analysis of the levels of Smad2 and phosphorylated Smad2 (Ser465/467) in mouse granulosa cells (mGC) treated with 50 ng/ml activin A (A) or infected with 50 MOI Ad-miR-181a (B) for 24 h. (C) mGC was infected with 50 MOI Ad-miR-181a for 24 h, and cells were then treated with 50 ng/ml activin A for another 24 h. The protein level of Smad2 and phosphorylated Smad2 were measured by Western blotting. (D) qRT-PCR analysis of Smad2 and cyclin D2 expression in mGC transfected with 50 nM siRNA duplexes targeting mouse Smad2 (siSmad2) or siRNA negative control for 48 h. *p

    Article Snippet: Immunoblotting was performed with primary antibodies against cyclin D2 (1∶1,000; Cell Signaling Technology, Danvers, MA, USA), phospho-Smad2 (Ser465/467; 1∶1,000; Cell Signaling Technology; also detects phosphorylated Smad3 at Ser423/425), Smad2 (1∶1,000; Cell Signaling Technology), PCNA (1∶500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), or acvr2a (1∶500; Santa Cruz Biotechnology). β-actin (1∶10,000; Abcam, Cambridge, MA, USA) was measured as an internal control.

    Techniques: Western Blot, Infection, Quantitative RT-PCR, Expressing, Transfection, Negative Control

    Stat3 inhibition induces growth arrest and cell death in human MM cell lines. ( A ) Immunoblot analysis of Stat3, pStat3-Y705, cyclin D1, cyclin D2 and survivin in human MM cell lines treated for 24 h with S3I-201 (RPMI-8226, 100 μM; H929 and EJM, 50 μM). β-actin levels are shown as loading control. ( B-D ) Quantification of cell growth and cell death of H929 ( B ), RPMI-8229 ( C ) and EJM ( D ) cells at various time points following treatment with S3I-201 at the indicated concentrations. Graphs are representative of two independent experiments.

    Journal: BMC Cancer

    Article Title: NF-?B2 mutation targets survival, proliferation and differentiation pathways in the pathogenesis of plasma cell tumors

    doi: 10.1186/1471-2407-12-203

    Figure Lengend Snippet: Stat3 inhibition induces growth arrest and cell death in human MM cell lines. ( A ) Immunoblot analysis of Stat3, pStat3-Y705, cyclin D1, cyclin D2 and survivin in human MM cell lines treated for 24 h with S3I-201 (RPMI-8226, 100 μM; H929 and EJM, 50 μM). β-actin levels are shown as loading control. ( B-D ) Quantification of cell growth and cell death of H929 ( B ), RPMI-8229 ( C ) and EJM ( D ) cells at various time points following treatment with S3I-201 at the indicated concentrations. Graphs are representative of two independent experiments.

    Article Snippet: Proteins (50 μg) were separated on SDS-polyacrylamide gels, transferred to nitrocellulose membranes, and probed with the following primary antibodies: mouse anti-cyclin D1 (sc-20044, 1:200, Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-cyclin D2 (#2924, 1:1000, Cell Signaling, Danvers, MA), rabbit anti-Stat3 (sc-482, 1:1000, Santa Cruz Biotechnology), rabbit anti-phospho-Stat3-Tyr705 (clone D3A7, #9145, 1:2000, Cell Signaling), mouse anti-survivin (clone D8, sc-17779, 1:100, Santa Cruz Biotechnology), and mouse anti-α-tubulin (B-5-1-2, 1:5000; Sigma-Aldrich).

    Techniques: Inhibition

    5-AZA blocked dexamethasone-induced downregulation of cyclin D2 protein and mRNA abundance in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and treated with dexamethasone (DEX) in the absence or presence of 5-aza-2'-deoxycytidine (5-AZA) for 48 hours. Cyclin D2 protein and mRNA abundance was determined by Western blot and quantitative real-time PCR, respectively. Data are mean ± S.E.M., n = 4 or 5, from different experiments. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Dexamethasone Induces Cardiomyocyte Terminal Differentiation via Epigenetic Repression of Cyclin D2 Gene

    doi: 10.1124/jpet.116.234104

    Figure Lengend Snippet: 5-AZA blocked dexamethasone-induced downregulation of cyclin D2 protein and mRNA abundance in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and treated with dexamethasone (DEX) in the absence or presence of 5-aza-2'-deoxycytidine (5-AZA) for 48 hours. Cyclin D2 protein and mRNA abundance was determined by Western blot and quantitative real-time PCR, respectively. Data are mean ± S.E.M., n = 4 or 5, from different experiments. * P

    Article Snippet: Membranes were incubated with anti-cyclin D2 (1:1000, Abcam) overnight at 4°C followed by washing.

    Techniques: Isolation, Western Blot, Real-time Polymerase Chain Reaction

    Dexamethasone downregulated cyclin D2 protein abundance in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and treated with dexamethasone (DEX) in the absence or presence of Ru486 for 48 hours. Cyclin D2 protein abundance was determined by Western blot. Data are mean ± S.E.M., n = 4 or 5, from different experiments. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Dexamethasone Induces Cardiomyocyte Terminal Differentiation via Epigenetic Repression of Cyclin D2 Gene

    doi: 10.1124/jpet.116.234104

    Figure Lengend Snippet: Dexamethasone downregulated cyclin D2 protein abundance in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and treated with dexamethasone (DEX) in the absence or presence of Ru486 for 48 hours. Cyclin D2 protein abundance was determined by Western blot. Data are mean ± S.E.M., n = 4 or 5, from different experiments. * P

    Article Snippet: Membranes were incubated with anti-cyclin D2 (1:1000, Abcam) overnight at 4°C followed by washing.

    Techniques: Isolation, Western Blot

    Overexpression of cyclin D2 in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and transfected with FlexiCcnD2 (+CcnD2) or Flexi empty construct (−CcnD4). Myocytes were then treated with dexamethasone (DEX) for 48 hours. Cyclin D2 protein abundance was determined by Western blot. STD, internal standard. Data are mean ± S.E.M., n = 4, from different experiments. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Dexamethasone Induces Cardiomyocyte Terminal Differentiation via Epigenetic Repression of Cyclin D2 Gene

    doi: 10.1124/jpet.116.234104

    Figure Lengend Snippet: Overexpression of cyclin D2 in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and transfected with FlexiCcnD2 (+CcnD2) or Flexi empty construct (−CcnD4). Myocytes were then treated with dexamethasone (DEX) for 48 hours. Cyclin D2 protein abundance was determined by Western blot. STD, internal standard. Data are mean ± S.E.M., n = 4, from different experiments. * P

    Article Snippet: Membranes were incubated with anti-cyclin D2 (1:1000, Abcam) overnight at 4°C followed by washing.

    Techniques: Over Expression, Isolation, Transfection, Construct, Western Blot

    Overexpression of cyclin D2 abrogated dexamethasone-induced terminal differentiation in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and transfected with FlexiCcnD2 (+CcnD2) or Flexi empty construct (−CcnD4). Myocytes were then treated with dexamethasone (DEX) for 48 hours. Cells were fixed and stained with Ki67 and α -actinin. The nuclei were stained with Hoechst. (A) Ki67 positive cardiomyocytes. (B) Binucleate cardiomyocytes. Data are mean ± S.E.M., n = 5 or 6. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Dexamethasone Induces Cardiomyocyte Terminal Differentiation via Epigenetic Repression of Cyclin D2 Gene

    doi: 10.1124/jpet.116.234104

    Figure Lengend Snippet: Overexpression of cyclin D2 abrogated dexamethasone-induced terminal differentiation in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and transfected with FlexiCcnD2 (+CcnD2) or Flexi empty construct (−CcnD4). Myocytes were then treated with dexamethasone (DEX) for 48 hours. Cells were fixed and stained with Ki67 and α -actinin. The nuclei were stained with Hoechst. (A) Ki67 positive cardiomyocytes. (B) Binucleate cardiomyocytes. Data are mean ± S.E.M., n = 5 or 6. * P

    Article Snippet: Membranes were incubated with anti-cyclin D2 (1:1000, Abcam) overnight at 4°C followed by washing.

    Techniques: Over Expression, Isolation, Transfection, Construct, Staining

    Dexamethasone increased cyclin D2 promoter methylation in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and treated with dexamethasone (DEX) in the absence or presence of 5-aza-2'-deoxycytidine (5-AZA) for 48 hours. Genomic DNA was extracted and 5-methylcytosine in CcnD2 proximal promoter was determined by MeDIP assays. Data are mean ± S.E.M., n = 4. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Dexamethasone Induces Cardiomyocyte Terminal Differentiation via Epigenetic Repression of Cyclin D2 Gene

    doi: 10.1124/jpet.116.234104

    Figure Lengend Snippet: Dexamethasone increased cyclin D2 promoter methylation in newborn cardiomyocytes. Cardiomyocytes were isolated from 2-day-old rats and treated with dexamethasone (DEX) in the absence or presence of 5-aza-2'-deoxycytidine (5-AZA) for 48 hours. Genomic DNA was extracted and 5-methylcytosine in CcnD2 proximal promoter was determined by MeDIP assays. Data are mean ± S.E.M., n = 4. * P

    Article Snippet: Membranes were incubated with anti-cyclin D2 (1:1000, Abcam) overnight at 4°C followed by washing.

    Techniques: Methylation, Isolation, Methylated DNA Immunoprecipitation

    Overexpression of cyclin D2 in H9C2 cells. H9C2 cells were transfected with either Flexi-empty (−CcnD2) or Flexi-CcnD2 (+CcnD2) constructs for 24 hours, followed by recovery for another 24 hours in fresh medium without transfection reagent. Transfection was performed in duplicate with one clone of Flexi-empty (Control) and two different clones of Flexi-CcnD2 (S and M). Whole cell lysates in RIPA buffer containing protease inhibitors were next made and protein concentration determined. Fifty micrograms total protein from each sample was electrophoresed by SDS-PAGE, followed by immunoblotting with cyclin D2 monoclonal antibody. Data are mean ± S.E.M., n = 4, from different experiments. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Dexamethasone Induces Cardiomyocyte Terminal Differentiation via Epigenetic Repression of Cyclin D2 Gene

    doi: 10.1124/jpet.116.234104

    Figure Lengend Snippet: Overexpression of cyclin D2 in H9C2 cells. H9C2 cells were transfected with either Flexi-empty (−CcnD2) or Flexi-CcnD2 (+CcnD2) constructs for 24 hours, followed by recovery for another 24 hours in fresh medium without transfection reagent. Transfection was performed in duplicate with one clone of Flexi-empty (Control) and two different clones of Flexi-CcnD2 (S and M). Whole cell lysates in RIPA buffer containing protease inhibitors were next made and protein concentration determined. Fifty micrograms total protein from each sample was electrophoresed by SDS-PAGE, followed by immunoblotting with cyclin D2 monoclonal antibody. Data are mean ± S.E.M., n = 4, from different experiments. * P

    Article Snippet: Membranes were incubated with anti-cyclin D2 (1:1000, Abcam) overnight at 4°C followed by washing.

    Techniques: Over Expression, Transfection, Construct, Clone Assay, Protein Concentration, SDS Page

    Recurrent mutations in CCND2 and MGA

    Journal: Nature genetics

    Article Title: The Genomic Landscape of Core-Binding Factor Acute Myeloid Leukemias

    doi: 10.1038/ng.3709

    Figure Lengend Snippet: Recurrent mutations in CCND2 and MGA

    Article Snippet: After transferring to PVDF membrane (iBlot, Life Technology), membranes were probed with total CCND2 (Abcam, ab3085) and GAPDH (Santa Cruz Biotechnology, 6C5) antibodies using the iBind system (Life Technology).

    Techniques:

    Knockdown of CITED1 upregulates the expression of the CKIs p21 and p27. a Western blotting analysis was performed to detect the expression levels of the cell cycle regulators cyclin A2, cyclin B1, cyclin D1, cyclin D2, cyclin D3, cyclin E1, cyclin E2, CDK2, CDK4, CDK6, p21 Cip1 , and p27 Kip1 in the indicated cells. α-tubulin was used as the loading control. b Silencing of CITED1 in the studied cells significantly inhibited the phosphorylation of pRb at the Ser608 residue. α-tubulin served as the sample loading control. c Silencing of CITED1 attenuated E2F transcriptional activity according to the E2F-luc reporter assay. The results are derived from three independent experiments and are expressed as the mean ± SD. *P

    Journal: Cell & Bioscience

    Article Title: CITED1 promotes proliferation of papillary thyroid cancer cells via the regulation of p21 and p27

    doi: 10.1186/s13578-018-0256-9

    Figure Lengend Snippet: Knockdown of CITED1 upregulates the expression of the CKIs p21 and p27. a Western blotting analysis was performed to detect the expression levels of the cell cycle regulators cyclin A2, cyclin B1, cyclin D1, cyclin D2, cyclin D3, cyclin E1, cyclin E2, CDK2, CDK4, CDK6, p21 Cip1 , and p27 Kip1 in the indicated cells. α-tubulin was used as the loading control. b Silencing of CITED1 in the studied cells significantly inhibited the phosphorylation of pRb at the Ser608 residue. α-tubulin served as the sample loading control. c Silencing of CITED1 attenuated E2F transcriptional activity according to the E2F-luc reporter assay. The results are derived from three independent experiments and are expressed as the mean ± SD. *P

    Article Snippet: The antibodies used for immunoblotting were as follows: anti-CITED1, anti-cyclin B1, anti-cyclin D3, anti-cyclin A2, anti-cyclin D1, anti-cyclin E1, anti-CDK4, anti-CDK6 (Abcam, Cambridge, MA), anti-cyclin D2, anti-CDK2 (BD Pharmingen, San Diego, CA), anti-cyclin E2, anti-p21Cip1, anti-p27Kip1 (Cell Signaling Technology, Beverly, MA), and anti-α-tubulin (Sigma-Aldrich, St. Louis, Missouri).

    Techniques: Expressing, Western Blot, Activity Assay, Reporter Assay, Derivative Assay

    Analysis of miR-15b/16-2 targets in miR-15b/16-2–deleted mouse B cells. ( A ) Western blot analysis of Cyclin D2 expression in B cells from knockout and wild-type mice after LPS stimulation at the indicated time points. β-Actin was used as a loading control. Fold change in protein expression is indicated. hs, hours. ( B ) Western blot analysis of Cyclin D2 and Cyclin D1 expression in B cells from knockout and wild-type mice. β-Actin was used as a loading control. ( C ) Relative quantification of Cyclin D1 and Cyclin D2 expression to the β-actin loading control in B cells from knockout and wild-type mice. ( D ) psiCHECK-2 vector with the CCND2 WT 3′ UTR insert and Mut miR-15b 3′ UTR (mut1, mut2, and mut3) containing a deletion of the miR-15b target site in the 3′ UTR was cotransfected with miR-15b or scrambled miR in 293HEK cells. Luciferase activity was recorded after 24 h. Data represent the mean ± SD from at least three independent experiments.

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

    Article Title: miR-15b/16-2 deletion promotes B-cell malignancies

    doi: 10.1073/pnas.1514954112

    Figure Lengend Snippet: Analysis of miR-15b/16-2 targets in miR-15b/16-2–deleted mouse B cells. ( A ) Western blot analysis of Cyclin D2 expression in B cells from knockout and wild-type mice after LPS stimulation at the indicated time points. β-Actin was used as a loading control. Fold change in protein expression is indicated. hs, hours. ( B ) Western blot analysis of Cyclin D2 and Cyclin D1 expression in B cells from knockout and wild-type mice. β-Actin was used as a loading control. ( C ) Relative quantification of Cyclin D1 and Cyclin D2 expression to the β-actin loading control in B cells from knockout and wild-type mice. ( D ) psiCHECK-2 vector with the CCND2 WT 3′ UTR insert and Mut miR-15b 3′ UTR (mut1, mut2, and mut3) containing a deletion of the miR-15b target site in the 3′ UTR was cotransfected with miR-15b or scrambled miR in 293HEK cells. Luciferase activity was recorded after 24 h. Data represent the mean ± SD from at least three independent experiments.

    Article Snippet: Western blot analysis was carried out using BCL2 (Santa Cruz Biotechnology), IGF1RB (Cell Signaling Technology), Cyclin D2 (BD Pharmingen), and GAPDH (GeneTex) antibodies.

    Techniques: Western Blot, Expressing, Knock-Out, Mouse Assay, Plasmid Preparation, Luciferase, Activity Assay

    miR-15b/16-2 expression in human CLL samples. ( A and B ) qRT-PCR analysis of miR-15b ( A ) and pri-miR-15b ( B ) in CLL samples. The number of patients analyzed and the P values are indicated. ( C and D ) Correlations between miR-15b and Cyclin D2 ( C ) or Cyclin D1 ( D ) expression were determined using the Spearman coefficient in normal and CLL samples (Cyclin D2: n = 50, P = 0.038; Cyclin D1: n = 50, P = 0.021). hsa, human. ( E and F ) ISH for miR-15b in human normal lymph node (LN) and CLL and DLBCL samples and quantification. ( G and H ) ISH for miR-15b ( H , Upper ) and miR-15b and Cyclin D1 ( H , Lower ) in human CLL, DLBCL, and MCL samples (asterisks indicate normal lymph node remnants showing moderate miR-15b expression). ( G ) Quantification of miR-15b and Cyclin D1 expression in CLL, DLBCL, and MCL samples. Error bars represent SD.

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

    Article Title: miR-15b/16-2 deletion promotes B-cell malignancies

    doi: 10.1073/pnas.1514954112

    Figure Lengend Snippet: miR-15b/16-2 expression in human CLL samples. ( A and B ) qRT-PCR analysis of miR-15b ( A ) and pri-miR-15b ( B ) in CLL samples. The number of patients analyzed and the P values are indicated. ( C and D ) Correlations between miR-15b and Cyclin D2 ( C ) or Cyclin D1 ( D ) expression were determined using the Spearman coefficient in normal and CLL samples (Cyclin D2: n = 50, P = 0.038; Cyclin D1: n = 50, P = 0.021). hsa, human. ( E and F ) ISH for miR-15b in human normal lymph node (LN) and CLL and DLBCL samples and quantification. ( G and H ) ISH for miR-15b ( H , Upper ) and miR-15b and Cyclin D1 ( H , Lower ) in human CLL, DLBCL, and MCL samples (asterisks indicate normal lymph node remnants showing moderate miR-15b expression). ( G ) Quantification of miR-15b and Cyclin D1 expression in CLL, DLBCL, and MCL samples. Error bars represent SD.

    Article Snippet: Western blot analysis was carried out using BCL2 (Santa Cruz Biotechnology), IGF1RB (Cell Signaling Technology), Cyclin D2 (BD Pharmingen), and GAPDH (GeneTex) antibodies.

    Techniques: Expressing, Quantitative RT-PCR, In Situ Hybridization

    miR-15b/16-2 expression in human CLL samples. ( A ) Correlations between miR-15b and Cyclin D2 ( Left ) or Cyclin D1 ( Right ) expression were determined using Spearman coefficient CLL samples from the TCGA database (Cyclin D2: P = 0.004; Cyclin D1: P = 0.027). ( B ) ISH for miR-15b in human normal lymph node and CLL and DLBCL samples (asterisks indicate normal lymph node remnants showing moderate miR-15b expression).

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

    Article Title: miR-15b/16-2 deletion promotes B-cell malignancies

    doi: 10.1073/pnas.1514954112

    Figure Lengend Snippet: miR-15b/16-2 expression in human CLL samples. ( A ) Correlations between miR-15b and Cyclin D2 ( Left ) or Cyclin D1 ( Right ) expression were determined using Spearman coefficient CLL samples from the TCGA database (Cyclin D2: P = 0.004; Cyclin D1: P = 0.027). ( B ) ISH for miR-15b in human normal lymph node and CLL and DLBCL samples (asterisks indicate normal lymph node remnants showing moderate miR-15b expression).

    Article Snippet: Western blot analysis was carried out using BCL2 (Santa Cruz Biotechnology), IGF1RB (Cell Signaling Technology), Cyclin D2 (BD Pharmingen), and GAPDH (GeneTex) antibodies.

    Techniques: Expressing, In Situ Hybridization

    Pilomatricomas express cyclins D1, D2, and D3. A: H E stain of pilomatricoma. Arrow indicates shadow cells; arrowhead indicates basoloid cells (×200), Tumor cells are focally positive for cyclin D1 ( B ), cyclin D2 ( C ), and cyclin D3 ( D ) ( arrows , ×200).

    Journal: The American Journal of Pathology

    Article Title: Differential Expression of Cyclin D1 in the Human Hair Follicle

    doi:

    Figure Lengend Snippet: Pilomatricomas express cyclins D1, D2, and D3. A: H E stain of pilomatricoma. Arrow indicates shadow cells; arrowhead indicates basoloid cells (×200), Tumor cells are focally positive for cyclin D1 ( B ), cyclin D2 ( C ), and cyclin D3 ( D ) ( arrows , ×200).

    Article Snippet: Takano Y, Kato Y, Masuda M, Ohshima Y, Okayasu I: Cyclin D2, but not cyclin D1, overexpression closely correlates with gastric cancer progression and prognosis.

    Techniques: Staining

    AZD1480 reduces the tumor growth of Kms.11 xenograft in mice associated with inhibition of JAK2/STAT3 and FGFR3 signaling and Cyclin D2 in vivo

    Journal: Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K

    Article Title: The novel JAK inhibitor AZD1480 blocks STAT3 and FGFR3 signaling, resulting in suppression of human myeloma cell growth and survival

    doi: 10.1038/leu.2010.289

    Figure Lengend Snippet: AZD1480 reduces the tumor growth of Kms.11 xenograft in mice associated with inhibition of JAK2/STAT3 and FGFR3 signaling and Cyclin D2 in vivo

    Article Snippet: Equal amounts of proteins were loaded onto a SDS-PAGE gel, transferred onto nitrocellulose membrane, and probed with the indicated antibody: rabbit polyclonal anti-phospho-JAK2 (Chemicon International); rabbit polyclonal anti-STAT3, anti-phospho-STAT3 (Tyr705), anti-phospho-STAT3 (Ser727), anti-p44/42 MAPK and anti-phospho-p44/42 MAPK (Cell Signaling Technology, Danvers, MA); mouse monoclonal anti-c-Myc, rabbit polyclonal anti-Mcl-1, rabbit polyclonal anti-Cyclin-D2 (Santa Cruz Biotechnology, Santa Cruz, CA); mouse monoclonal Bcl-xL antibody (Calbiochem); and mouse monoclonal β-actin antibody (Sigma-Aldrich, St Louis, MO).

    Techniques: Mouse Assay, Inhibition, In Vivo

    AZD1480 inhibits the IL-6-inducible upregulation of the STAT3 target genes c-Myc and Cyclin-D2

    Journal: Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K

    Article Title: The novel JAK inhibitor AZD1480 blocks STAT3 and FGFR3 signaling, resulting in suppression of human myeloma cell growth and survival

    doi: 10.1038/leu.2010.289

    Figure Lengend Snippet: AZD1480 inhibits the IL-6-inducible upregulation of the STAT3 target genes c-Myc and Cyclin-D2

    Article Snippet: Equal amounts of proteins were loaded onto a SDS-PAGE gel, transferred onto nitrocellulose membrane, and probed with the indicated antibody: rabbit polyclonal anti-phospho-JAK2 (Chemicon International); rabbit polyclonal anti-STAT3, anti-phospho-STAT3 (Tyr705), anti-phospho-STAT3 (Ser727), anti-p44/42 MAPK and anti-phospho-p44/42 MAPK (Cell Signaling Technology, Danvers, MA); mouse monoclonal anti-c-Myc, rabbit polyclonal anti-Mcl-1, rabbit polyclonal anti-Cyclin-D2 (Santa Cruz Biotechnology, Santa Cruz, CA); mouse monoclonal Bcl-xL antibody (Calbiochem); and mouse monoclonal β-actin antibody (Sigma-Aldrich, St Louis, MO).

    Techniques:

    RNAi-mediated knockdown of cyclin D3 induces G 2 /M arrest and polyploidy. (A) MLE cells were transfected with either scrambled RNA or cyclin D2 siRNA or cyclin D3 siRNA. 72 h later, cells were harvested; lysates were resolved on SDS-PAGE prior to cyclin

    Journal: Cell Cycle

    Article Title: FBXL2 is a ubiquitin E3 ligase subunit that triggers mitotic arrest

    doi: 10.4161/cc.10.20.17742

    Figure Lengend Snippet: RNAi-mediated knockdown of cyclin D3 induces G 2 /M arrest and polyploidy. (A) MLE cells were transfected with either scrambled RNA or cyclin D2 siRNA or cyclin D3 siRNA. 72 h later, cells were harvested; lysates were resolved on SDS-PAGE prior to cyclin

    Article Snippet: Mouse monoclonal cyclin D2, cyclin D3 and CDK11 antibodies and were from Abcam.

    Techniques: Transfection, SDS Page

    Overexpression of FBXL2 induces G 2 /M arrest by selectively degrading cyclin D2 and cyclin D3. (A) FACS analysis of A549 cells transfected with either empty vector or FBXL2 plasmid; viable cells were quantified and graphed (n = 3). (B) FACS analysis of

    Journal: Cell Cycle

    Article Title: FBXL2 is a ubiquitin E3 ligase subunit that triggers mitotic arrest

    doi: 10.4161/cc.10.20.17742

    Figure Lengend Snippet: Overexpression of FBXL2 induces G 2 /M arrest by selectively degrading cyclin D2 and cyclin D3. (A) FACS analysis of A549 cells transfected with either empty vector or FBXL2 plasmid; viable cells were quantified and graphed (n = 3). (B) FACS analysis of

    Article Snippet: Mouse monoclonal cyclin D2, cyclin D3 and CDK11 antibodies and were from Abcam.

    Techniques: Over Expression, FACS, Transfection, Plasmid Preparation