ccug 44156 sweden pos ccug 55226 belgium pos ccug 44258 sweden pos ccug 48515 sweden pos trichomonas vaginalis atcc 30001 nd pos  (ATCC)

Journal: Cell reports

Article Title: Nuclear translocation of an aminoacyl-tRNA synthetase may mediate a chronic “integrated stress response”

doi: 10.1016/j.celrep.2023.112632

Figure Lengend Snippet:

Article Snippet: shE2F1 , Santa Cruz Biotechnology , Catalog #: sc-29297-SH.

Techniques: Recombinant, Extraction, Silver Staining, CCK-8 Assay, Software

Journal: The World Journal of Men's Health

Article Title: Docetaxel Enhances Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand-Mediated Apoptosis in Prostate Cancer Cells via Epigenetic Gene Regulation by Enhancer of Zeste Homolog 2

doi: 10.5534/wjmh.220073

Figure Lengend Snippet: Expressions of EZH2 and E2F1 in DU 145 cells. WB assay shows decreases of EZH2 and E2F1 expressions after docetaxel treatment (A). Using knockdown of E2F1, the expressions of E2F1 and EZH2 are decreased (B), and TRAIL (100 ng/mL) treatment causes significant cell death in siE2F-1 DU 145 cells ( *** p<0.001) (C). DR4: death receptor 4, DR5: death receptor 5, EZH2: enhancer of zeste homolog 2, ns: not significant, TRAIL: tumor necrosis factor-related apoptosis-inducing ligand, WB: western blot.

Article Snippet: Small interfering RNA (siRNA) against human E2F1 (siE2F1) (sc-29297) and control siRNA (scRNA) (sc-37007) were purchased from Santa Cruz Biotechnology.

Techniques: Knockdown, Western Blot

Journal: PLoS ONE

Article Title: E2F-1 Directly Regulates Thrombospondin 1 Expression

doi: 10.1371/journal.pone.0013442

Figure Lengend Snippet: A, schematic depiction of six TSP1 promoter deletion constructs in the region −2033 to +750bp. The transcription initial site is designated as +1. B, quantification of E2F-1 transcriptional activity on six TSP1 promoter constructs. 293 cells were used for assays and 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. C, dose response of E2F1 on TSP1 promoter activity in U2OS cells. D, the expression of different doses of Flag-tagged E2F1 was verified by Western blot using anti-flag monoclonal antibody, the amounts of Flag-E2F-1 for transfection were indicated. E, the promoter luciferase reporters of E2F1 targeted gene ARF , Apaf1 and CyclinE were used as controls to evaluate the transcriptional activity of E2F-1 on TSP1 promoter, 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. F, schematic depiction of four TSP1 promoter deletion constructs in the region −413 to 0 bp. G, quantification of E2F-1 transcriptional activity on four TSP1 promoter constructs, 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. H, the expression of HA-tagged E2F-1 was verified by Western blot using anti-HA monoclonal antibody.

Article Snippet: U2OS cells growing 6-well plate were used for verifying E2F-1 knockdown by E2F-1 siRNA using a monoclonal antibody against E2F-1(Santa Cruz).

Techniques: Construct, Activity Assay, Transfection, Expressing, Western Blot, Luciferase

Journal: PLoS ONE

Article Title: E2F-1 Directly Regulates Thrombospondin 1 Expression

doi: 10.1371/journal.pone.0013442

Figure Lengend Snippet: A, the partial sequence of TSP1 promoter (−393 to +27bp). Three potential E2F-1 binding sites were marked by boxes and substitution mutations in those sites were indicated underline. B, schematic of eight TSP1 promoter mutation constructs in the region −393 to 0bp. The three potential E2F-1 binding sites were marked by dark circles and mutations were marked by “X”. C, quantification of E2F-1 transcriptional activity on eight TSP1 promoter mutation constructs, 0.2 µg Flag tagged E2F-1/per well was used for transfection in 24-well plates. D, schematic depiction of E2F-1 DNA binding region (amino acid 110–194) cloned into pVP16 vector. E, quantification of E2F-1 transcriptional activity on three TSP1 promoter mutants, 0.2 µg VP-16E2F-1(110–194) or VP-16/per well was used for transfection in 24-well plates. F, quantification of wild-type E2F-1 and E2F-1(E132) transcriptional activity on TSP1 promoter (−2033–0) luciferase reporter, 0.2 µg vector/per well was used for transfection. G, the expression of HA-tagged wild-type E2F-1 and E2F-1 DNA binding site mutant (E132) was verified by Western blot using anti-Flag monoclonal antibody. H, quantification of pRB influence on E2F-1 transcriptional activity on TSP1 promoter (−2033–0) luciferase reporter construct, the expression of HA tagged human pRB was verified by Western blot and the amounts of HA-pRB for transfection were indicated. I, quantification of E2F-1 deletion mutants on TSP1 promoter (−2033–0) luciferase reporter construct. J, the expression of flag-tagged E2F-1 deletion constructs was verified by Western blot using anti-Flag monoclonal antibody, 0.2 µg different construct/per well was used for transfection. K, quantification of E2F-1 siRNA on TSP1 promoter (−2033–0) luciferase reporter activity. L, the knockdown of E2F1 by siRNA in U2OS cells was verified by Western blot using anti-E2F1 monoclonal antibody, a siRNA targeting GFP was used as control, 0.6 µg siRNA expression vector/per well targeting E2F-1 or GFP was used for transfection in 6-well plates.

Article Snippet: U2OS cells growing 6-well plate were used for verifying E2F-1 knockdown by E2F-1 siRNA using a monoclonal antibody against E2F-1(Santa Cruz).

Techniques: Sequencing, Binding Assay, Mutagenesis, Construct, Activity Assay, Transfection, Clone Assay, Plasmid Preparation, Luciferase, Expressing, Western Blot

Journal: PLoS ONE

Article Title: E2F-1 Directly Regulates Thrombospondin 1 Expression

doi: 10.1371/journal.pone.0013442

Figure Lengend Snippet: A, schematic diagram depicts the region of the TSP1 and actin genes that were amplified. The positions of PCR primers used to detect TSP1 and actin promoter fragments are indicated by arrows. B, 293 cells were treated with formaldehyde to create cross-links between E2F-1 and chromatin. The chromatin was isolated, sheared, and immunoprecipitated (IP) using monoclonal antibody against human E2F-1, or mouse IgG as control. The presence of chromatin fragments corresponding to the TSP1 gene or to the β-actin gene promoter was assessed by PCR using gene-specific primers. The gel shows the recovery of TSP1 and actin gene fragments from the protein-chromatin input on the lane 1 (from left to right) as well as those recovered after immunoprecipitation with the anti-E2F-1 antibody (lane 2, up), and with mouse IgG (land 3).

Article Snippet: U2OS cells growing 6-well plate were used for verifying E2F-1 knockdown by E2F-1 siRNA using a monoclonal antibody against E2F-1(Santa Cruz).

Techniques: Amplification, Isolation, Immunoprecipitation

Journal: PLoS ONE

Article Title: E2F-1 Directly Regulates Thrombospondin 1 Expression

doi: 10.1371/journal.pone.0013442

Figure Lengend Snippet: Northern blot analysis of TSP1 expression in 293 cells transfected with HA empty vector of HA-E2F-1 expression vector. β-actin was used as internal control.

Article Snippet: U2OS cells growing 6-well plate were used for verifying E2F-1 knockdown by E2F-1 siRNA using a monoclonal antibody against E2F-1(Santa Cruz).

Techniques: Northern Blot, Expressing, Transfection, Plasmid Preparation

Journal: PLoS ONE

Article Title: E2F-1 Directly Regulates Thrombospondin 1 Expression

doi: 10.1371/journal.pone.0013442

Figure Lengend Snippet: A, schematic depiction of six TSP1 promoter deletion constructs in the region −2033 to +750bp. The transcription initial site is designated as +1. B, quantification of E2F-1 transcriptional activity on six TSP1 promoter constructs. 293 cells were used for assays and 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. C, dose response of E2F1 on TSP1 promoter activity in U2OS cells. D, the expression of different doses of Flag-tagged E2F1 was verified by Western blot using anti-flag monoclonal antibody, the amounts of Flag-E2F-1 for transfection were indicated. E, the promoter luciferase reporters of E2F1 targeted gene ARF , Apaf1 and CyclinE were used as controls to evaluate the transcriptional activity of E2F-1 on TSP1 promoter, 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. F, schematic depiction of four TSP1 promoter deletion constructs in the region −413 to 0 bp. G, quantification of E2F-1 transcriptional activity on four TSP1 promoter constructs, 0.2 µg HA tagged E2F-1/per well was used for transfection in 24-well plates. H, the expression of HA-tagged E2F-1 was verified by Western blot using anti-HA monoclonal antibody.

Article Snippet: U2OS cells growing 6-well plate were used for verifying E2F-1 knockdown by E2F-1 siRNA using a monoclonal antibody against E2F-1(Santa Cruz).

Techniques: Construct, Activity Assay, Transfection, Expressing, Western Blot, Luciferase

Journal: PLoS ONE

Article Title: E2F-1 Directly Regulates Thrombospondin 1 Expression

doi: 10.1371/journal.pone.0013442

Figure Lengend Snippet: A, the partial sequence of TSP1 promoter (−393 to +27bp). Three potential E2F-1 binding sites were marked by boxes and substitution mutations in those sites were indicated underline. B, schematic of eight TSP1 promoter mutation constructs in the region −393 to 0bp. The three potential E2F-1 binding sites were marked by dark circles and mutations were marked by “X”. C, quantification of E2F-1 transcriptional activity on eight TSP1 promoter mutation constructs, 0.2 µg Flag tagged E2F-1/per well was used for transfection in 24-well plates. D, schematic depiction of E2F-1 DNA binding region (amino acid 110–194) cloned into pVP16 vector. E, quantification of E2F-1 transcriptional activity on three TSP1 promoter mutants, 0.2 µg VP-16E2F-1(110–194) or VP-16/per well was used for transfection in 24-well plates. F, quantification of wild-type E2F-1 and E2F-1(E132) transcriptional activity on TSP1 promoter (−2033–0) luciferase reporter, 0.2 µg vector/per well was used for transfection. G, the expression of HA-tagged wild-type E2F-1 and E2F-1 DNA binding site mutant (E132) was verified by Western blot using anti-Flag monoclonal antibody. H, quantification of pRB influence on E2F-1 transcriptional activity on TSP1 promoter (−2033–0) luciferase reporter construct, the expression of HA tagged human pRB was verified by Western blot and the amounts of HA-pRB for transfection were indicated. I, quantification of E2F-1 deletion mutants on TSP1 promoter (−2033–0) luciferase reporter construct. J, the expression of flag-tagged E2F-1 deletion constructs was verified by Western blot using anti-Flag monoclonal antibody, 0.2 µg different construct/per well was used for transfection. K, quantification of E2F-1 siRNA on TSP1 promoter (−2033–0) luciferase reporter activity. L, the knockdown of E2F1 by siRNA in U2OS cells was verified by Western blot using anti-E2F1 monoclonal antibody, a siRNA targeting GFP was used as control, 0.6 µg siRNA expression vector/per well targeting E2F-1 or GFP was used for transfection in 6-well plates.

Article Snippet: U2OS cells growing 6-well plate were used for verifying E2F-1 knockdown by E2F-1 siRNA using a monoclonal antibody against E2F-1(Santa Cruz).

Techniques: Sequencing, Binding Assay, Mutagenesis, Construct, Activity Assay, Transfection, Clone Assay, Plasmid Preparation, Luciferase, Expressing, Western Blot

Journal: PLoS ONE

Article Title: E2F-1 Directly Regulates Thrombospondin 1 Expression

doi: 10.1371/journal.pone.0013442

Figure Lengend Snippet: A, schematic diagram depicts the region of the TSP1 and actin genes that were amplified. The positions of PCR primers used to detect TSP1 and actin promoter fragments are indicated by arrows. B, 293 cells were treated with formaldehyde to create cross-links between E2F-1 and chromatin. The chromatin was isolated, sheared, and immunoprecipitated (IP) using monoclonal antibody against human E2F-1, or mouse IgG as control. The presence of chromatin fragments corresponding to the TSP1 gene or to the β-actin gene promoter was assessed by PCR using gene-specific primers. The gel shows the recovery of TSP1 and actin gene fragments from the protein-chromatin input on the lane 1 (from left to right) as well as those recovered after immunoprecipitation with the anti-E2F-1 antibody (lane 2, up), and with mouse IgG (land 3).

Article Snippet: U2OS cells growing 6-well plate were used for verifying E2F-1 knockdown by E2F-1 siRNA using a monoclonal antibody against E2F-1(Santa Cruz).

Techniques: Amplification, Isolation, Immunoprecipitation

Journal: PLoS ONE

Article Title: E2F-1 Directly Regulates Thrombospondin 1 Expression

doi: 10.1371/journal.pone.0013442

Figure Lengend Snippet: Northern blot analysis of TSP1 expression in 293 cells transfected with HA empty vector of HA-E2F-1 expression vector. β-actin was used as internal control.

Article Snippet: U2OS cells growing 6-well plate were used for verifying E2F-1 knockdown by E2F-1 siRNA using a monoclonal antibody against E2F-1(Santa Cruz).

Techniques: Northern Blot, Expressing, Transfection, Plasmid Preparation

Journal: Cell reports

Article Title: E2F1 transcription factor mediates a link between fat and islets to promote β cell proliferation in response to acute insulin resistance

doi: 10.1016/j.celrep.2022.111436

Figure Lengend Snippet: The expression of E2F family genes in islets from S961-treated mice versus islets from vehicle-treated mice

Article Snippet: Lentiviral particles for murine E2F1 short hairpin RNA (shRNA) (sc-29297-V) and control scramble shRNA (sc-108080) were purchased from Santa Cruz.

Techniques: Expressing

Journal: Cell reports

Article Title: E2F1 transcription factor mediates a link between fat and islets to promote β cell proliferation in response to acute insulin resistance

doi: 10.1016/j.celrep.2022.111436

Figure Lengend Snippet: (A and C–F) The sera were collected from mice at day 7 of treatment. β cells, mouse islets, or human islets were treated with 20% serum from S-961- or Ns-treated C57BL/6J male mice. (A) MTT assay (absorbance at 570 nm) in control, IRS1KO, IRS2KO, and βIRKO β cells. *p < 0.05, **p < 0.01 (n = 6 biological replicates). (B) Western blot of indicated proteins in scramble- or E2F1-knockdown control β cells. (C) MTT assay in scramble- or E2F1-knockdown control β cells. *p < 0.05, **p < 0.01 (n = 6 biological replicates). (D) Left: representative images of indicated β cells. Nuclei are stained blue, and EdU+ nuclei are stained red. Right: number of ErdU+ β cells. *p < 0.05, **p < 0.01 (n = 6 biological replicates). (E) Mouse islets were incubated with 20% serum from indicated mice in the presence or absence of HLM006474 for 24 h. Left: representative images of islets. Insulin is stained red, nuclei are stained blue, and EdU+ nuclei are stained green. Yellow arrowheads indicate insulin+ and EdU+ cells. Right: number of ErdU+ β cells in the islets. *p < 0.05 (n = 5 mice/group). (F) Human islets from non-diabetes donors were incubated with 20% serum from indicated mice in the presence or absence of HLM006474 for 48 h. Yellow arrowheads indicate insulin+ and EdU+ cells. Left: representative images of islets. Insulin is stained red, nuclei are stained blue, and EdU+ nuclei are stained green. Right: number of ErdU+ β cells in the islets. *p < 0.05 (n = 7 donors). (A and C–F) All data from three or more independent experiments are represented as mean ± SEM. An unpaired two-tailed Student’s t test (A and C), a one-way ANOVA (D and E), and a Mann-Whitney U test (F) were performed. (B) Data are representative of three independent experiments.

Article Snippet: Lentiviral particles for murine E2F1 short hairpin RNA (shRNA) (sc-29297-V) and control scramble shRNA (sc-108080) were purchased from Santa Cruz.

Techniques: MTT Assay, Control, Western Blot, Knockdown, Staining, Incubation, Two Tailed Test, MANN-WHITNEY

Journal: Cell reports

Article Title: E2F1 transcription factor mediates a link between fat and islets to promote β cell proliferation in response to acute insulin resistance

doi: 10.1016/j.celrep.2022.111436

Figure Lengend Snippet:

Article Snippet: Lentiviral particles for murine E2F1 short hairpin RNA (shRNA) (sc-29297-V) and control scramble shRNA (sc-108080) were purchased from Santa Cruz.

Techniques: Virus, Control, shRNA, Recombinant, Protein Extraction, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Reverse Transcription, SYBR Green Assay, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Proliferation Assay, Imaging, Microarray, Knock-Out, Software