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kristian helin  (Addgene inc)


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    Addgene inc kristian helin
    Kristian Helin, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/kristian+helin/pCMVHA-E2F5+(Plasmid+%2324213)/pm41644695-265-52-54
    Average 92 stars, based on 7 article reviews
    kristian helin - by Bioz Stars, 2026-09
    92/100 stars

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    Plasmid Preparation:

    Article Title: E2F1 combined with LINC01004 super-enhancer to promote hepatocellular carcinoma cell proliferation and metastasis.
    Article Snippet: The human hepatocellular carcinoma cells (HepG2 and SKHep-1 cell lines) were maintained at 37 °C in humidified 5% CO2 atmosphere containing 5% CO2 in Dulbecco’s modified Eagle medium (Hyclone, Logan, USA) supplemented with 10% fetal bovine serum (FBS) (CLARK, Richmond, USA). .. Cells were plated and grown until they were reached 70–80% confluent. pCMVHA E2F1 was a gift from Kristian Helin [23] (Addgene plasmid # 24,225, https:// www. addge ne. org/ 24225/, RRID:Addgene_24225). pLV hUbCdCas9 KRAB-T2A-GFP was a gift from Charles Gersbach (Addgene plasmid # 67,620, https:// www. addge ne. org/ 67620/, RRID:Addgene_67620) [24]. .. Then plasmids and siRNAs (GenePharma, Suzhou, China) were transfected using lipofectamine 3000 (Invitrogen, Carlsbad, USA). si-NC: sence 5′- UUC UCC GAA CGU GUC ACG UTT-3′, antisence 5′- ACG UGA CAC GUU CGG AGA ATT-3′; si-LINC01004-1: sence 5′- GGU UCA AGG UAU AAG CUA AACTT-3′; antisence 5′- UUA GCU UAU ACC UUG AAC CUATT-3′; si-LINC01004-2: sence 5′- GGG AAU AUG UUG UGU UCU AAUTT-3′; antisence 5′- UAG AAC ACA ACA UAU UCC CGATT-3′;

    Article Title: Suppression of canonical TGF-β signaling enables GATA4 to interact with H3K27me3 demethylase JMJD3 to promote cardiomyogenesis.
    Article Snippet: .. pBabe-X GATA4, Hand2, Mef2c, Tbx5, miR-1, and miR-133 were generated previously [14,15]. pCMV-HA-JMJD3 was a gift from Kristian Helin (Addgene plasmid #24167 [63]). pBabe-X-HA-UTX was subcloned from pCMV-HA-UTX, a gift from Kristian Helin (Addgene plasmid #24168 [63]). pBabe-X-HA-SMAD2 was subcloned from pCMV5B-HASMAD2, a gift from Jeff Wrana (Addgene plasmid #11734 [64]). pBabe-puro-SMAD7-HA was a gift from Sam Thiagalingam (Addgene plasmid #37044 [65]). .. pCMV-HA-JMJD3 was a gift from Kristian Helin (Addgene plasmid #24167 [63]).

    Article Title: McIdas localizes to centrioles and controls centriole numbers through PLK4-dependent phosphorylation
    Article Snippet: Final expression vectors were generated by LR recombination using the Gateway LR clonase II enzyme mix (Invitrogen, 11791) between attL-containing entry clones and attR-containing pLVDest-CAG destination vectors to produce the constructs: pLVDest GFP-McIdas (mouse), pLVDest GFP-McIdas (human), pLVDest GFP-McIdas NES1mut, pLVDest GFP-McIdas NES2mut, pLVDest GFP-McIdas NES1/2mut, pLVDest GFP-McIdas 10 A, pLVDest GFP-McIdas 10D, pLVDest GFP-McIdas 4A_N, pLVDest GFP-McIdas 3A_cc and pLVDest GFP-McIdas 3A_C. .. Additional plasmids used in this study include: pcDNA3.1 McIdas-HA and pcDNA3.1 HA control vectors (Pefani et al, ), pCMV HA-STIL (Ohta et al, ), pEBTet HsSAS6-GFP and pEBTet HsSAS6_SNAP-FLAG, kindly provided by Dr. Pierre Gonczy (1 μg/ml tetracycline was used for efficient induction (Keller et al, ) and pCMVHA-E2F5, a gift from Kristian Helin (Addgene plasmid #24213). .. U2OS (ATCC, HTB-96), HeLa (ATCC, CCL-2) and 293 T (ATCC, CRL-3216) cells were cultured in DMEM (Gibco) supplemented with 10% fetal bovine serum (FBS) in 5% CO 2 at 37 °C. hTERT RPE-1 cells (ATCC, CRL-4000) were cultured in DMEM/F12 (Sigma-Aldrich) supplemented with 10% FBS under the same conditions.

    Article Title: Alternative mRNA Splicing Controls the Functions of the Histone H3K27 Demethylase UTX/KDM6A.
    Article Snippet: The UTX long isoform cDNA was purchased from Genscript (CloneID OHu24601). .. The cDNA for UTX lacking exon 14 sequences was PCR amplified from pCMV-HA-UTX, which was a gift from Kristian Helin (Addgene plasmid #24168) [8]. .. All UTX isoforms and mutants were generated via PCR cloning into the pDONR221 vector or via site-directed mutagenesis using the QuikChange strategy (Agilent, Santa Clara, CA, USA).

    Article Title: The Role of Crosstalk between AR3 and E2F1 in Drug Resistance in Prostate Cancer Cells.
    Article Snippet: Auranofin (BML-EI206-0100) was purchased from Enzo Life Sciences (Farmingdale, NY, USA) and dissolved in DMSO (stock concentration: 200 mg/mL). .. The plasmids used in the present study are as follows: the E2F1 expression vector pCMVHA E2F1 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24225) [18], the E2F1 DNA binding mutant vector pCMV E2F1 E132 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24224) [19], the E2F1 promoter luciferase vector pGL2-AN was a gift from William Kaelin (Addgene, Cambridge, MA, USA, plasmid # 20950) [20], and the AR expression vector and ARR2-Luc vector were generated as previously described [21,22]. .. The antibodies used in this study include: Rabbit polyclonal anti-AR3 [22], Anti-AR3(Clone RM7; ReMab, San Francisco, CA, USA), Anti-AR-FL (sc-815; Santa Cruz, Dallas, TX, USA), Anti-AR (sc-816; Santa Cruz, Dallas, TX, USA), Anti-α-tubulin (clone DM1A; Sigma, St Louis, MO, USA), Anti-E2F1 (clones KH20 and KH95; Millipore, Burlington, MA, USA), Anti-E2F1 (A300-766A; Sigma, St Louis, MO, USA), Anti-Rb (9309S; CST, Danvers, MA, USA), Anti-PARP1(sc-8007; Santa Cruz, Dallas, TX, USA), Anti-Caspase3 (9661S; CST, Danvers, MA, USA), Anti-GAPDH (sc-365062; Santa Cruz, Dallas, TX, USA), and Anti-Ki67 (9449; CST, Danvers, MA, USA).

    Article Title: Nox4 promotes endothelial differentiation through chromatin remodeling.
    Article Snippet: .. The transfection mixture was then added dropwise and after 4 h the medium was changed to Hek293 growth medium. pCMV-HA-JMJD3 was a gift from Kristian Helin (Addgene plasmid # 24167; http://n2t.net/addgene:24167; RRID: Addgene_24167). ..

    other:

    Article Title: McIdas localizes to centrioles and controls centriole numbers through PLK4-dependent phosphorylation.
    Article Snippet: Final expression vectors were generated by LR recombination using the Gateway LR clonase II enzyme mix (Invitrogen, 11791) between attL-containing entry clones and attR-containing pLVDest-CAG destination vectors to produce the constructs: pLVDest GFP-McIdas (mouse), pLVDest GFP-McIdas (human), pLVDest GFP-McIdas NES1mut, pLVDest GFP-McIdas NES2mut, pLVDest GFP-McIdas NES1/2mut, pLVDest GFP-McIdas 10 A, pLVDest GFP-McIdas 10D, pLVDest GFP-McIdas 4A_N, pLVDest GFP-McIdas 3A_cc and pLVDest GFP-McIdas 3A_C.

    Generated:

    Article Title: Suppression of canonical TGF-β signaling enables GATA4 to interact with H3K27me3 demethylase JMJD3 to promote cardiomyogenesis.
    Article Snippet: .. pBabe-X GATA4, Hand2, Mef2c, Tbx5, miR-1, and miR-133 were generated previously [14,15]. pCMV-HA-JMJD3 was a gift from Kristian Helin (Addgene plasmid #24167 [63]). pBabe-X-HA-UTX was subcloned from pCMV-HA-UTX, a gift from Kristian Helin (Addgene plasmid #24168 [63]). pBabe-X-HA-SMAD2 was subcloned from pCMV5B-HASMAD2, a gift from Jeff Wrana (Addgene plasmid #11734 [64]). pBabe-puro-SMAD7-HA was a gift from Sam Thiagalingam (Addgene plasmid #37044 [65]). .. pCMV-HA-JMJD3 was a gift from Kristian Helin (Addgene plasmid #24167 [63]).

    Article Title: The Role of Crosstalk between AR3 and E2F1 in Drug Resistance in Prostate Cancer Cells.
    Article Snippet: Auranofin (BML-EI206-0100) was purchased from Enzo Life Sciences (Farmingdale, NY, USA) and dissolved in DMSO (stock concentration: 200 mg/mL). .. The plasmids used in the present study are as follows: the E2F1 expression vector pCMVHA E2F1 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24225) [18], the E2F1 DNA binding mutant vector pCMV E2F1 E132 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24224) [19], the E2F1 promoter luciferase vector pGL2-AN was a gift from William Kaelin (Addgene, Cambridge, MA, USA, plasmid # 20950) [20], and the AR expression vector and ARR2-Luc vector were generated as previously described [21,22]. .. The antibodies used in this study include: Rabbit polyclonal anti-AR3 [22], Anti-AR3(Clone RM7; ReMab, San Francisco, CA, USA), Anti-AR-FL (sc-815; Santa Cruz, Dallas, TX, USA), Anti-AR (sc-816; Santa Cruz, Dallas, TX, USA), Anti-α-tubulin (clone DM1A; Sigma, St Louis, MO, USA), Anti-E2F1 (clones KH20 and KH95; Millipore, Burlington, MA, USA), Anti-E2F1 (A300-766A; Sigma, St Louis, MO, USA), Anti-Rb (9309S; CST, Danvers, MA, USA), Anti-PARP1(sc-8007; Santa Cruz, Dallas, TX, USA), Anti-Caspase3 (9661S; CST, Danvers, MA, USA), Anti-GAPDH (sc-365062; Santa Cruz, Dallas, TX, USA), and Anti-Ki67 (9449; CST, Danvers, MA, USA).

    Control:

    Article Title: McIdas localizes to centrioles and controls centriole numbers through PLK4-dependent phosphorylation
    Article Snippet: Final expression vectors were generated by LR recombination using the Gateway LR clonase II enzyme mix (Invitrogen, 11791) between attL-containing entry clones and attR-containing pLVDest-CAG destination vectors to produce the constructs: pLVDest GFP-McIdas (mouse), pLVDest GFP-McIdas (human), pLVDest GFP-McIdas NES1mut, pLVDest GFP-McIdas NES2mut, pLVDest GFP-McIdas NES1/2mut, pLVDest GFP-McIdas 10 A, pLVDest GFP-McIdas 10D, pLVDest GFP-McIdas 4A_N, pLVDest GFP-McIdas 3A_cc and pLVDest GFP-McIdas 3A_C. .. Additional plasmids used in this study include: pcDNA3.1 McIdas-HA and pcDNA3.1 HA control vectors (Pefani et al, ), pCMV HA-STIL (Ohta et al, ), pEBTet HsSAS6-GFP and pEBTet HsSAS6_SNAP-FLAG, kindly provided by Dr. Pierre Gonczy (1 μg/ml tetracycline was used for efficient induction (Keller et al, ) and pCMVHA-E2F5, a gift from Kristian Helin (Addgene plasmid #24213). .. U2OS (ATCC, HTB-96), HeLa (ATCC, CCL-2) and 293 T (ATCC, CRL-3216) cells were cultured in DMEM (Gibco) supplemented with 10% fetal bovine serum (FBS) in 5% CO 2 at 37 °C. hTERT RPE-1 cells (ATCC, CRL-4000) were cultured in DMEM/F12 (Sigma-Aldrich) supplemented with 10% FBS under the same conditions.

    Polymerase Chain Reaction:

    Article Title: Alternative mRNA Splicing Controls the Functions of the Histone H3K27 Demethylase UTX/KDM6A.
    Article Snippet: The UTX long isoform cDNA was purchased from Genscript (CloneID OHu24601). .. The cDNA for UTX lacking exon 14 sequences was PCR amplified from pCMV-HA-UTX, which was a gift from Kristian Helin (Addgene plasmid #24168) [8]. .. All UTX isoforms and mutants were generated via PCR cloning into the pDONR221 vector or via site-directed mutagenesis using the QuikChange strategy (Agilent, Santa Clara, CA, USA).

    Amplification:

    Article Title: Alternative mRNA Splicing Controls the Functions of the Histone H3K27 Demethylase UTX/KDM6A.
    Article Snippet: The UTX long isoform cDNA was purchased from Genscript (CloneID OHu24601). .. The cDNA for UTX lacking exon 14 sequences was PCR amplified from pCMV-HA-UTX, which was a gift from Kristian Helin (Addgene plasmid #24168) [8]. .. All UTX isoforms and mutants were generated via PCR cloning into the pDONR221 vector or via site-directed mutagenesis using the QuikChange strategy (Agilent, Santa Clara, CA, USA).

    Expressing:

    Article Title: The Role of Crosstalk between AR3 and E2F1 in Drug Resistance in Prostate Cancer Cells.
    Article Snippet: Auranofin (BML-EI206-0100) was purchased from Enzo Life Sciences (Farmingdale, NY, USA) and dissolved in DMSO (stock concentration: 200 mg/mL). .. The plasmids used in the present study are as follows: the E2F1 expression vector pCMVHA E2F1 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24225) [18], the E2F1 DNA binding mutant vector pCMV E2F1 E132 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24224) [19], the E2F1 promoter luciferase vector pGL2-AN was a gift from William Kaelin (Addgene, Cambridge, MA, USA, plasmid # 20950) [20], and the AR expression vector and ARR2-Luc vector were generated as previously described [21,22]. .. The antibodies used in this study include: Rabbit polyclonal anti-AR3 [22], Anti-AR3(Clone RM7; ReMab, San Francisco, CA, USA), Anti-AR-FL (sc-815; Santa Cruz, Dallas, TX, USA), Anti-AR (sc-816; Santa Cruz, Dallas, TX, USA), Anti-α-tubulin (clone DM1A; Sigma, St Louis, MO, USA), Anti-E2F1 (clones KH20 and KH95; Millipore, Burlington, MA, USA), Anti-E2F1 (A300-766A; Sigma, St Louis, MO, USA), Anti-Rb (9309S; CST, Danvers, MA, USA), Anti-PARP1(sc-8007; Santa Cruz, Dallas, TX, USA), Anti-Caspase3 (9661S; CST, Danvers, MA, USA), Anti-GAPDH (sc-365062; Santa Cruz, Dallas, TX, USA), and Anti-Ki67 (9449; CST, Danvers, MA, USA).

    Binding Assay:

    Article Title: The Role of Crosstalk between AR3 and E2F1 in Drug Resistance in Prostate Cancer Cells.
    Article Snippet: Auranofin (BML-EI206-0100) was purchased from Enzo Life Sciences (Farmingdale, NY, USA) and dissolved in DMSO (stock concentration: 200 mg/mL). .. The plasmids used in the present study are as follows: the E2F1 expression vector pCMVHA E2F1 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24225) [18], the E2F1 DNA binding mutant vector pCMV E2F1 E132 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24224) [19], the E2F1 promoter luciferase vector pGL2-AN was a gift from William Kaelin (Addgene, Cambridge, MA, USA, plasmid # 20950) [20], and the AR expression vector and ARR2-Luc vector were generated as previously described [21,22]. .. The antibodies used in this study include: Rabbit polyclonal anti-AR3 [22], Anti-AR3(Clone RM7; ReMab, San Francisco, CA, USA), Anti-AR-FL (sc-815; Santa Cruz, Dallas, TX, USA), Anti-AR (sc-816; Santa Cruz, Dallas, TX, USA), Anti-α-tubulin (clone DM1A; Sigma, St Louis, MO, USA), Anti-E2F1 (clones KH20 and KH95; Millipore, Burlington, MA, USA), Anti-E2F1 (A300-766A; Sigma, St Louis, MO, USA), Anti-Rb (9309S; CST, Danvers, MA, USA), Anti-PARP1(sc-8007; Santa Cruz, Dallas, TX, USA), Anti-Caspase3 (9661S; CST, Danvers, MA, USA), Anti-GAPDH (sc-365062; Santa Cruz, Dallas, TX, USA), and Anti-Ki67 (9449; CST, Danvers, MA, USA).

    Mutagenesis:

    Article Title: The Role of Crosstalk between AR3 and E2F1 in Drug Resistance in Prostate Cancer Cells.
    Article Snippet: Auranofin (BML-EI206-0100) was purchased from Enzo Life Sciences (Farmingdale, NY, USA) and dissolved in DMSO (stock concentration: 200 mg/mL). .. The plasmids used in the present study are as follows: the E2F1 expression vector pCMVHA E2F1 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24225) [18], the E2F1 DNA binding mutant vector pCMV E2F1 E132 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24224) [19], the E2F1 promoter luciferase vector pGL2-AN was a gift from William Kaelin (Addgene, Cambridge, MA, USA, plasmid # 20950) [20], and the AR expression vector and ARR2-Luc vector were generated as previously described [21,22]. .. The antibodies used in this study include: Rabbit polyclonal anti-AR3 [22], Anti-AR3(Clone RM7; ReMab, San Francisco, CA, USA), Anti-AR-FL (sc-815; Santa Cruz, Dallas, TX, USA), Anti-AR (sc-816; Santa Cruz, Dallas, TX, USA), Anti-α-tubulin (clone DM1A; Sigma, St Louis, MO, USA), Anti-E2F1 (clones KH20 and KH95; Millipore, Burlington, MA, USA), Anti-E2F1 (A300-766A; Sigma, St Louis, MO, USA), Anti-Rb (9309S; CST, Danvers, MA, USA), Anti-PARP1(sc-8007; Santa Cruz, Dallas, TX, USA), Anti-Caspase3 (9661S; CST, Danvers, MA, USA), Anti-GAPDH (sc-365062; Santa Cruz, Dallas, TX, USA), and Anti-Ki67 (9449; CST, Danvers, MA, USA).

    Luciferase:

    Article Title: The Role of Crosstalk between AR3 and E2F1 in Drug Resistance in Prostate Cancer Cells.
    Article Snippet: Auranofin (BML-EI206-0100) was purchased from Enzo Life Sciences (Farmingdale, NY, USA) and dissolved in DMSO (stock concentration: 200 mg/mL). .. The plasmids used in the present study are as follows: the E2F1 expression vector pCMVHA E2F1 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24225) [18], the E2F1 DNA binding mutant vector pCMV E2F1 E132 was a gift from Kristian Helin (Addgene, Cambridge, MA, USA, plasmid # 24224) [19], the E2F1 promoter luciferase vector pGL2-AN was a gift from William Kaelin (Addgene, Cambridge, MA, USA, plasmid # 20950) [20], and the AR expression vector and ARR2-Luc vector were generated as previously described [21,22]. .. The antibodies used in this study include: Rabbit polyclonal anti-AR3 [22], Anti-AR3(Clone RM7; ReMab, San Francisco, CA, USA), Anti-AR-FL (sc-815; Santa Cruz, Dallas, TX, USA), Anti-AR (sc-816; Santa Cruz, Dallas, TX, USA), Anti-α-tubulin (clone DM1A; Sigma, St Louis, MO, USA), Anti-E2F1 (clones KH20 and KH95; Millipore, Burlington, MA, USA), Anti-E2F1 (A300-766A; Sigma, St Louis, MO, USA), Anti-Rb (9309S; CST, Danvers, MA, USA), Anti-PARP1(sc-8007; Santa Cruz, Dallas, TX, USA), Anti-Caspase3 (9661S; CST, Danvers, MA, USA), Anti-GAPDH (sc-365062; Santa Cruz, Dallas, TX, USA), and Anti-Ki67 (9449; CST, Danvers, MA, USA).

    Transfection:

    Article Title: Nox4 promotes endothelial differentiation through chromatin remodeling.
    Article Snippet: .. The transfection mixture was then added dropwise and after 4 h the medium was changed to Hek293 growth medium. pCMV-HA-JMJD3 was a gift from Kristian Helin (Addgene plasmid # 24167; http://n2t.net/addgene:24167; RRID: Addgene_24167). ..



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