Journal:

Article Title: Delayed replication timing leads to delayed mitotic chromosome condensation and chromosomal instability of chromosome translocations

doi: 10.1073/pnas.241355098

Figure Lengend Snippet: Delay of mitotic chromosome condensation in tumor cell lines. (A) Mitotic chromosome spreads were scored for the presence of chromosomes with DMC in tumor derived cells. Chromosomes with DMC were scored in mitotic spreads harvested either in the presence (+) or absence (−) of colcemid from the blood of two normal individuals (46XY and 46XX) and from the tumor-derived cell lines RH30, CRL-5845, CRL-5824, HTB-118, WERI-RB1, and HTB-81. Following propidium iodine staining, a minimum of 100 mitotic spreads was scored for each sample. Sequential G-banding (C) and FISH (B) with a chromosome 3 paint on a mitotic spread from RH30 cells prepared in the absence of colcemid. This probe hybridized to a nonrearranged chromosome 3 (arrow), and to a chromosome with DMC (✻ and arrow). (D and E) Mitotic chromosome spreads showing two copies of the der(1q) (D) that hybridize to a chromosome 1 paint (E). (F) Mitotic spreads were analyzed sequentially by G-banding (G) followed by FISH with a chromosome 1 paint (1). The DNA was visualized in the FISH analyses by staining with propidium iodide.

Article Snippet: C2C12, CRL-5845, CRL-5824, HTB-81, HTB-118, WERI-RB1, and HELA cells were from the American Type Culture Collection.

Techniques: Derivative Assay, Staining

Journal: mBio

Article Title: The Mus musculus Papillomavirus Type 1 E7 Protein Binds to the Retinoblastoma Tumor Suppressor: Implications for Viral Pathogenesis

doi: 10.1128/mBio.02277-21

Figure Lengend Snippet: MmuPV1 E7 binds but does not destabilize the retinoblastoma tumor suppressor protein, RB1. Alignment of the N-terminal sequences of MmuPV1 E7 and the canine papillomavirus 2 (CPV2), γ1-HPV4, γ24-HPV197, β1-HPV8, and α9-HPV16. Identical residues are marked by black boxes, and chemically similar residues are shaded in gray. The position of the LXCXE RB1 binding site is highlighted. (A) A cartoon of the domain structure of E7 is shown on top, with the N-terminal sequences that show similarity the conserved regions 1 and 2 (CR1, CR2) of adenovirus E1A proteins indicated. MmuPV1 E7 (M E7) can interact with RB1 by immunoprecipitation/immunoblot analysis. (B) Duplicate cultures of HCT116 cells were transfected with an N-terminally HA/FLAG epitope-tagged MmuPV1 E7 expression plasmid, and coprecipitated RB1 protein was detected by immunoblotting. MmuPV1 E7 (M E7) can interact with murine Rb1 by immunoprecipitation/immunoblot analysis. (C) NIH 3T3 murine fibroblasts were transfected with an N-terminally HA/FLAG epitope-tagged MmuPV1 E7 expression plasmid and coprecipitated RB1 protein detected by immunoblotting. MmuPV1 E7 does not destabilize RB. SAOS-2 human osteosarcoma cells that do not express detectable endogenous RB1 were transfected with an RB1 expression vector and increasing amounts of an expression vector for N-terminally HA/FLAG epitope-tagged MmuPV1 followed by Western blotting to assess RB1 steady levels by Western blotting. GFP was cotransfected and assessed by Western blotting to control for transfection efficiency. HPV16 E7 (16 E7) was used as a positive control. (D) A representative blot from one of four experiments is shown.

Article Snippet: The RB1 truncation plasmids pSG5-HA-Rb 1–928, 1–792, and 379–928 were kind gifts from Bill Sellers (Broad Institute).

Techniques: Binding Assay, Immunoprecipitation, Western Blot, Transfection, FLAG-tag, Expressing, Plasmid Preparation, Control, Positive Control

Journal: mBio

Article Title: The Mus musculus Papillomavirus Type 1 E7 Protein Binds to the Retinoblastoma Tumor Suppressor: Implications for Viral Pathogenesis

doi: 10.1128/mBio.02277-21

Figure Lengend Snippet: LXCXE binding cleft in the RB1 protein is not necessary for MmuPV1 E7 binding. SAOS-2 human osteosarcoma cells were transfected with expression vectors for wild-type RB1 or the RB1 L mutant that contains three amino acid mutations in the LXCXE binding cleft in combination with MmuPV1 or HPV16 E7 expression vectors. MmuPV1 E7 binds wild type and RB1 L with similar efficiency, whereas HPV16 E7 binds and causes degradation of RB1 but not RB1 L . The results shown are representative of two independent experiments. A cartoon of the RB1 L mutant is shown at the top.

Article Snippet: The RB1 truncation plasmids pSG5-HA-Rb 1–928, 1–792, and 379–928 were kind gifts from Bill Sellers (Broad Institute).

Techniques: Binding Assay, Transfection, Expressing, Mutagenesis

Journal: mBio

Article Title: The Mus musculus Papillomavirus Type 1 E7 Protein Binds to the Retinoblastoma Tumor Suppressor: Implications for Viral Pathogenesis

doi: 10.1128/mBio.02277-21

Figure Lengend Snippet: Disruption of the LXCXE binding cleft in RB1 does not influence MmuPV1’s ability to cause papillomas in vivo . Sites on the ears of both wild-type and RB1 L mutant FVB/N mice were scarified and infected with 10 8 VGE of MmuPV1. Mice were treated with 300 mJ UVB the next day and then monitored for papilloma formation over 4 months. (A) MmuPV1 induced warts in wild-type and RB1 L mutant FVB/N mice with a similar incidence (Fisher’s exact test, P = 1, two-sided). Warts arising in wild-type and RB1 L mutant FVB/N mice share similar microscopic features. (B) Warts from both mouse genotypes were harvested, serially sectioned, and stained with hematoxylin and eosin (H&E), processed to detect Ki67 (Ki67, red; DAPI, blue) by immunofluorescence, and MCM7 by immunohistochemistry.

Article Snippet: The RB1 truncation plasmids pSG5-HA-Rb 1–928, 1–792, and 379–928 were kind gifts from Bill Sellers (Broad Institute).

Techniques: Disruption, Binding Assay, In Vivo, Mutagenesis, Infection, Staining, Immunofluorescence, Immunohistochemistry

Journal: mBio

Article Title: The Mus musculus Papillomavirus Type 1 E7 Protein Binds to the Retinoblastoma Tumor Suppressor: Implications for Viral Pathogenesis

doi: 10.1128/mBio.02277-21

Figure Lengend Snippet: C-terminal domain of RB1 is necessary for MmuPV1 E7 binding. (A) Schematic representation of the RB1 protein and the expression plasmids used. HA-epitope tagged versions of full-length RB1 and the two truncation mutants 1–792 and 379–928 were expressed in SAOS-2 human osteosarcoma cells in combination with FLAG/HA epitope-tagged MmuPV1 (M E7) and HPV16 E7 (16 E7) expression vectors. (B) After immunoprecipitation with FLAG antibodies, HA-tagged E7 and coprecipitated RB1 proteins were detected by HA immunoblotting. The result shown is representative of two independent experiments.

Article Snippet: The RB1 truncation plasmids pSG5-HA-Rb 1–928, 1–792, and 379–928 were kind gifts from Bill Sellers (Broad Institute).

Techniques: Binding Assay, Expressing, Immunoprecipitation, Western Blot

Journal: mBio

Article Title: The Mus musculus Papillomavirus Type 1 E7 Protein Binds to the Retinoblastoma Tumor Suppressor: Implications for Viral Pathogenesis

doi: 10.1128/mBio.02277-21

Figure Lengend Snippet: The RB1 binding site maps to the MmuPV1 E7 C terminus. Sequence alignment of the C-terminal domains of MmuPV1, canine papillomavirus 2 (CPV2), γ1-HPV4, γ24-HPV197, β1-HPV8, and α9-HPV16 E7. Identical residues are marked by black boxes, and chemically similar residues are shaded in gray. The position of the CXXC motifs that form a zinc-binding site is shown. (A) The position of the aspartate residue at position 90 (D90) that is important for RB1 binding is indicated by a red box. (B) Various FLAG/HA-tagged MmuPV1 E7 mutants were expressed in HCT116 cells, and coprecipitated RB1 was detected by immunoblotting. The result shown is representative of six independent experiments. (C) Immunoprecipitation Western blot analyses to assess RB1 binding by various MmuPV1 E7 D90 mutants. The result shown is representative of two independent experiments. Immunoprecipitation/Western blot analysis documenting that MmuPV1 E7 D90A is defective for binding to murine Rb1. (D) Wild-type MmuPV1 E7 and the D90A mutant were transiently expressed in NIH 3T3 cells and binding assessed by immunoprecipitation/Western blotting. The blot shown is representative of 2 independent experiments. Quantifications of E7-coprecipitated RB1 or Rb1 are normalized to the amount of E7 that is precipitated and are shown underneath.

Article Snippet: The RB1 truncation plasmids pSG5-HA-Rb 1–928, 1–792, and 379–928 were kind gifts from Bill Sellers (Broad Institute).

Techniques: Binding Assay, Sequencing, Residue, Western Blot, Immunoprecipitation, Mutagenesis

Journal: Journal of Ginseng Research

Article Title: New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

doi: 10.5142/jgr.2012.36.2.205

Figure Lengend Snippet: Representative HPLC chromatogram of mixed standards (A), red ginseng powder (B), and red ginseng extract (C). 1, Rg 1 (15.53 min); 2, Re (16.12 min); 3, Rf (32.66 min); 4, Rh 1 (37.67 min); 5, Rg 2 (S) (38.04 min); 6, Rg 2 (R) (39.22 min); 7, Rb 1 (40.89 min); 8, Rc (42.74 min); 9, Rb 2 (44.44 min); 10, Rd (46.86 min); 11, Rg 3 (S) (56.15 min); 12, Rg 3 (R) (56.80 min).

Article Snippet: Ginsenoside Rg 1 , Re, Rf, Rh 1 , Rb 1 , Rc, Rb 2 , Rd, Rg 3 (S), and Rg 3 (R) were purchased from Chromadex Co. (Irvine, CA, USA) and ginsenoside Rg 2 (S) and Rg 2 (R) were obtained from Embo Lab. (Seoul, Korea).

Techniques:

Journal: Journal of Ginseng Research

Article Title: New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

doi: 10.5142/jgr.2012.36.2.205

Figure Lengend Snippet: Linearity of calibration curve for ginsenosides

Article Snippet: Ginsenoside Rg 1 , Re, Rf, Rh 1 , Rb 1 , Rc, Rb 2 , Rd, Rg 3 (S), and Rg 3 (R) were purchased from Chromadex Co. (Irvine, CA, USA) and ginsenoside Rg 2 (S) and Rg 2 (R) were obtained from Embo Lab. (Seoul, Korea).

Techniques:

Journal: Journal of Ginseng Research

Article Title: New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

doi: 10.5142/jgr.2012.36.2.205

Figure Lengend Snippet: Ginsenosides amount (mg/g) and precision result using red ginseng powder (means of 9 determinations over 3 concentration levels)

Article Snippet: Ginsenoside Rg 1 , Re, Rf, Rh 1 , Rb 1 , Rc, Rb 2 , Rd, Rg 3 (S), and Rg 3 (R) were purchased from Chromadex Co. (Irvine, CA, USA) and ginsenoside Rg 2 (S) and Rg 2 (R) were obtained from Embo Lab. (Seoul, Korea).

Techniques: Concentration Assay

Journal: Journal of Ginseng Research

Article Title: New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

doi: 10.5142/jgr.2012.36.2.205

Figure Lengend Snippet: Recovery and RSD values of red ginseng powder (9 determinations over 3 concentration levels)

Article Snippet: Ginsenoside Rg 1 , Re, Rf, Rh 1 , Rb 1 , Rc, Rb 2 , Rd, Rg 3 (S), and Rg 3 (R) were purchased from Chromadex Co. (Irvine, CA, USA) and ginsenoside Rg 2 (S) and Rg 2 (R) were obtained from Embo Lab. (Seoul, Korea).

Techniques: Concentration Assay

Journal: Journal of Ginseng Research

Article Title: New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

doi: 10.5142/jgr.2012.36.2.205

Figure Lengend Snippet: Ginsenosides amount (mg/g) and precision result using red ginseng extract (means of 9 determinations over 3 concentration levels)

Article Snippet: Ginsenoside Rg 1 , Re, Rf, Rh 1 , Rb 1 , Rc, Rb 2 , Rd, Rg 3 (S), and Rg 3 (R) were purchased from Chromadex Co. (Irvine, CA, USA) and ginsenoside Rg 2 (S) and Rg 2 (R) were obtained from Embo Lab. (Seoul, Korea).

Techniques: Concentration Assay

Journal: Journal of Ginseng Research

Article Title: New Method for Simultaneous Quantification of 12 Ginsenosides in Red Ginseng Powder and Extract: In-house Method Validation

doi: 10.5142/jgr.2012.36.2.205

Figure Lengend Snippet: Recovery and RSD values of red ginseng extract (9 determinations over 3 concentration levels)

Article Snippet: Ginsenoside Rg 1 , Re, Rf, Rh 1 , Rb 1 , Rc, Rb 2 , Rd, Rg 3 (S), and Rg 3 (R) were purchased from Chromadex Co. (Irvine, CA, USA) and ginsenoside Rg 2 (S) and Rg 2 (R) were obtained from Embo Lab. (Seoul, Korea).

Techniques: Concentration Assay

Journal: Pharmacology Research & Perspectives

Article Title: Intestinal, portal, and peripheral profiles of daikenchuto (TU-100)'s active ingredients after oral administration

doi: 10.1002/prp2.165

Figure Lengend Snippet: Time-dependent changes in intraluminal concentrations of ginseng ingredients Rb1 (left panels) and Rg1 (right panels) in various loci of rat small intestine. A small intestine was cut into four pieces (#1-4, from oral to anal side) at the indicated times after oral administration of TU-100 (1 g/kg BW), and the luminal concentrations of ingredients were determined as described in Materials and Methods. The percentage of the amount of each ingredient detected in various loci relative to that contained in the dose of administered TU-100 was calculated. Values represent means ± SD, n = 3. Closed columns represent values after β -glucuronidase treatment. Open columns represent values without treatment. Rb1, ginsenoside Rb 1 ; Rg1, ginsenoside Rg 1 ; TU-100, daikenchuto.

Article Snippet: The following authentic standards and internal standards were isolated or synthesized by Tsumura & Co.: hydroxy- α -sanshool (HAS), hydroxy- β -sanshool (HBS), γ -sanshool (GS), [6]-gingerol (6G), [6]-shogaol (6S), [10]-gingerol (10G), [10]- shogaol (10S), ginsenoside Rg 1 (Rg1), ginsenoside Rb 1 (Rb1), and schizandrin.

Techniques:

Journal: Journal of Ginseng Research

Article Title: Antiviral Effect of Korean Red Ginseng Extract and Ginsenosides on Murine Norovirus and Feline Calicivirus as Surrogates for Human Norovirus

doi: 10.5142/jgr.2011.35.4.429

Figure Lengend Snippet: Effect of purified ginsenoside Rg 1 . (A) Titration of feline calicivirus on Crandell Reese feline kidney cells pretreated with purified ginsenoside Rg 1 for 24 h. (B) Titration of murine norovirus on RAW264.7 cells pretreated with purified ginsenoside Rg 1 for 24 h. Values are mean±SD. The symbol * is used to indicate statistical significance with p <0.01. TCID 50 , tissue culture infectious dose.

Article Snippet: Purified ginsenoside Rb 1 (catalog no. ASB-00007191-005) and Rg 1 (catalog no. ASB-00007221-005) were purchased from Chromadex (Irvine, CA, USA) and re-dissolved into distilled water

Techniques: Purification, Titration

Journal: Journal of Ginseng Research

Article Title: Antiviral Effect of Korean Red Ginseng Extract and Ginsenosides on Murine Norovirus and Feline Calicivirus as Surrogates for Human Norovirus

doi: 10.5142/jgr.2011.35.4.429

Figure Lengend Snippet: Effect of purified ginsenoside Rb 1 . (A) Titration of feline calicivirus on Crandell Reese feline kidney cells pretreated with purified ginsenoside Rb 1 for 24 h. (B) Titration of murine norovirus on RAW264.7 cells pretreated with purified ginsenoside Rb 1 for 24 h. Values are mean±SD. The symbol * is used to indicate statistical significance with p <0.01. TCID 50 , tissue culture infectious dose.

Article Snippet: Purified ginsenoside Rb 1 (catalog no. ASB-00007191-005) and Rg 1 (catalog no. ASB-00007221-005) were purchased from Chromadex (Irvine, CA, USA) and re-dissolved into distilled water

Techniques: Purification, Titration