Journal:

Article Title: On the Intraoperative Molecular Status of Renal Allografts after Vascular Reperfusion and Clinical Outcomes

doi: 10.1681/ASN.2005020210

Figure Lengend Snippet: List of studied genes

Article Snippet: The intraoperative immunosuppressive regimen consisted of 1.5 mg/kg of thymoglobulin (Sangstat, Fremont, CA) given in a slow infusion begun before the transplant procedure or 20 mg of anti-CD25 mAb (Simulect; Novartis, East Hanover, NJ) and solumedrol 500 mg intravenously.

Techniques:

Journal:

Article Title: On the Intraoperative Molecular Status of Renal Allografts after Vascular Reperfusion and Clinical Outcomes

doi: 10.1681/ASN.2005020210

Figure Lengend Snippet: Prediction of clinical outcomes

Article Snippet: The intraoperative immunosuppressive regimen consisted of 1.5 mg/kg of thymoglobulin (Sangstat, Fremont, CA) given in a slow infusion begun before the transplant procedure or 20 mg of anti-CD25 mAb (Simulect; Novartis, East Hanover, NJ) and solumedrol 500 mg intravenously.

Techniques:

Journal: EMBO Molecular Medicine

Article Title: Increased CSF Aβ during the very early phase of cerebral Aβ deposition in mouse models

doi: 10.15252/emmm.201505026

Figure Lengend Snippet: Human Aβ exhibits a biphasic profile in CSF of APP transgenic mice A, B Aβ40 and Aβ42 concentrations in CSF of male APP23 mice (heterozygous; 3 ( n = 14), 6 ( n = 12), 8 ( n = 10), 12 ( n = 11), 19 ( n = 9), and 25 ( n = 6) months of age). CSF Aβ40 ( F (1, 56) = 22.351, P < 0.001) as well as CSF Aβ42 ( F (1, 56) = 38.597, P < 0.001) followed a significant quadratic trend. C Aβ42/40 ratio in CSF of APP23 mice showed a delayed decrease with age ( F (1, 56) = 53.894, P < 0.001). D, E Aβ40 and Aβ42 concentrations in CSF of male and female APP24 mice (homozygous; 2 ( n = 13), 3–4 ( n = 16), 7–8 ( n = 15), 18–19 ( n = 14), 24 ( n = 16), and 30 ( n = 18) months of age). CSF Aβ40 followed a significant quadratic trend ( F (1, 86) = 6.678, P = 0.011) and CSF Aβ42 best fitted a cubic trend ( F (1, 86) = 30.599, P < 0.001). F Aβ42/40 ratio in CSF of APP24 mice showed a delayed decrease with age ( F (1, 86) = 64.936, P < 0.001). G, H Aβ40 and Aβ42 in the CSF of female APP51 mice (heterozygous; 3 ( n = 6), 15 ( n = 8), and 24 ( n = 8) months of age; 22 mice in total). CSF Aβ40 ( F (1, 19) = 37.349, P < 0.001) as well as CSF Aβ42 ( F (1, 19) = 107.670, P < 0.001) followed a significant quadratic trend. I Aβ42/40 ratio in CSF of APP51 mice showed a delayed decrease with age ( F (1, 19) = 26.367, P < 0.001). Data information: Post hoc Dunnett's test was employed for group comparisons, which were always conducted between the youngest group and all other groups. (Observed CSF Aβ40 or Aβ42 changes were independent of batch.) All data are represented as group means ± SEM; * P < 0.05; ** P < 0.01; and *** P < 0.001.

Article Snippet: Male and female 2- to 30-month-old homozygous APP24 mice (Abramowski et al , ) were bred at both the Novartis Mouse facility (Basel, Switzerland) and the Hertie Institute for Clinical Brain Research (Tübingen, Germany).

Techniques: Transgenic Assay

Journal: EMBO Molecular Medicine

Article Title: Increased CSF Aβ during the very early phase of cerebral Aβ deposition in mouse models

doi: 10.15252/emmm.201505026

Figure Lengend Snippet: Human Aβ in CSF and brain of APP transgenic mice A APP23 CSF Aβ40 and Aβ42 in the same animals as shown in Fig . CSF Aβ42 and Aβ40 are expressed as percentages of levels measured in the youngest age group. B, C Aβ40 and Aβ42 (pmol/g wet brain) in the FA-soluble brain extract from the same APP23 mice showed a robust increase with age; ANOVA revealed a significant cubic trend ( F (1, 56) = 221.114, P < 0.001 and F (1, 56) = 370.947, P < 0.001, respectively). D APP24 CSF Aβ40 and Aβ42 in the same animals shown in Fig as percentage of the youngest age group. E, F Aβ40 and Aβ42 (pmol/g wet brain) in the brain from the same APP24 mice also showed a robust increase with age; ANOVA revealed a significant cubic trend ( F (1, 86) = 202.173, P < 0.001 and F (1, 86) = 139.941, P < 0.001, respectively). G APP51 CSF Aβ40 and Aβ42 in the same animals shown in Fig as percentages of levels in the youngest age group. H, I Aβ40 and Aβ42 (pmol/g wet brain) in the brain from the same APP51 mice showed a robust increase with age; ANOVA revealed a significant quadratic trend ( F (1, 19) = 12.960, P = 0.002 and F (1, 19) = 19.366, P < 0.001, respectively). Data information: Post hoc Dunnett's test group comparisons were always conducted between the youngest group and all other groups. All data are represented as group means ± SEM; * P < 0.05; ** P < 0.01; and *** P < 0.001.

Article Snippet: Male and female 2- to 30-month-old homozygous APP24 mice (Abramowski et al , ) were bred at both the Novartis Mouse facility (Basel, Switzerland) and the Hertie Institute for Clinical Brain Research (Tübingen, Germany).

Techniques: Transgenic Assay

Journal: EMBO Molecular Medicine

Article Title: Increased CSF Aβ during the very early phase of cerebral Aβ deposition in mouse models

doi: 10.15252/emmm.201505026

Figure Lengend Snippet: Brain sAPPβ shows an age-related increase in APP23, APP24, and APP51 mice sAPPβ was measured in Triton X-100 brain extracts from largely the same mice as analyzed in Figs and and is expressed as percentages of levels measured in the youngest age group. Swedish sAPPβ showed an age-dependent increase in APP23 mice following a linear trend ( F (1, 83) = 52.914, P < 0.001); APP23 from two independent batches were included in this analysis (see Materials and Methods and Supplementary Fig for details). Swedish sAPPβ showed an age-dependent increase in APP24 mice following a linear trend ( F (1, 84) = 11.264, P = 0.001). Human wild-type sAPPβ showed an age-dependent increase in APP51 following a quadratic trend ( F (1, 18) = 68.980, P < 0.001). Data information: Post hoc Dunnett's test group comparisons were always conducted between the youngest group and all other groups. All data are represented as group means ± SEM; * P < 0.05; ** P < 0.01; and *** P < 0.001. For absolute values, see Supplementary Fig .

Article Snippet: Male and female 2- to 30-month-old homozygous APP24 mice (Abramowski et al , ) were bred at both the Novartis Mouse facility (Basel, Switzerland) and the Hertie Institute for Clinical Brain Research (Tübingen, Germany).

Techniques:

Journal: Translational Oncology

Article Title: Anti-Angiogenic/Vascular Effects of the mTOR Inhibitor Everolimus Are Not Detectable by FDG/FLT-PET 1

doi:

Figure Lengend Snippet: Everolimus decreases FDG uptake by human H596 lung tumor xenografts. Human tumors were created by s.c. implantation of viable H596 tumor tissue in Harlan athymic mice. (A) For efficacy, mice were treated with vehicle or with everolimus (10 mg/kg p.o.) for 21 days. Results show mean ± SEM. (B and C) A separate cohort of tumor-bearing mice was studied by FDG-PET as described in the Materials and Methods section, immediately before and 2 days after treatment with everolimus. Results show the individual SUVs for tumors (n = 10 per treatment group) and the mean ± SEM fractional effect of treatment, where *P < .05, **P <.01. (D) Histology and IHC of ablated tumors on day 2 after completion of the second PET scan.

Article Snippet: Female Brown-Norway rats and C57BL/6 mice were obtained from Charles River (France) and female Harlan Hsd:Npa nu/nu (nude) athymic mice were obtained from the Novartis breeding stock (Basel, Switzerland).

Techniques:

Journal: Translational Oncology

Article Title: Anti-Angiogenic/Vascular Effects of the mTOR Inhibitor Everolimus Are Not Detectable by FDG/FLT-PET 1

doi:

Figure Lengend Snippet: Everolimus inhibits growth of insensitive human tumors but does not affect FDG uptake. Human HCT116 (colon) and KB31 (cervical) were created by s.c. injection of cells in Harlan athymic mice. In all cases, mice were treated daily p.o. with vehicle or everolimus (10 mg/kg). (A and B) Efficacy in HCT116 tumors for 1 week (A) or 2 weeks (B) showing the mean ± SEM with the associated T/CTVol at the end point. (C and D) FDG-PET in HCT116 tumors in the respective experiment showing the mean fractional change for each treatment compared with day 0 (C) or the individual tumor SUV at two different time points (D). (E and F) Efficacy in KB31 tumors with the fractional change in FDG-PET compared with day 0 for each treatment, where *P < .05, ***P < .001.

Article Snippet: Female Brown-Norway rats and C57BL/6 mice were obtained from Charles River (France) and female Harlan Hsd:Npa nu/nu (nude) athymic mice were obtained from the Novartis breeding stock (Basel, Switzerland).

Techniques: Injection

Journal: Translational Oncology

Article Title: Anti-Angiogenic/Vascular Effects of the mTOR Inhibitor Everolimus Are Not Detectable by FDG/FLT-PET 1

doi:

Figure Lengend Snippet: Everolimus decreases FLT uptake by human H596 lung tumor xenografts. Human tumors were created by s.c. implantation of viable H596 tumor tissue in Harlan athymic mice. Tumors were studied by FLT-PET as described in the Materials and Methods section, immediately before and 2 days after treatment with everolimus (10 mg/kg p.o.). (A) Representative experiment showing horizontal whole-body images of mice with tumors (blue arrow) before and after treatment with everolimus; red arrow indicates the bladder. (B) Individual SUVs for tumors (n = 6 per treatment group), and the mean ± SEM fractional effect of treatment, where **P < .01. (C) Histology and IHC of ablated tumors on day 2 after completion of the second PET scan.

Article Snippet: Female Brown-Norway rats and C57BL/6 mice were obtained from Charles River (France) and female Harlan Hsd:Npa nu/nu (nude) athymic mice were obtained from the Novartis breeding stock (Basel, Switzerland).

Techniques:

Journal: Translational Oncology

Article Title: Anti-Angiogenic/Vascular Effects of the mTOR Inhibitor Everolimus Are Not Detectable by FDG/FLT-PET 1

doi:

Figure Lengend Snippet: Everolimus does not affect FLT uptake by human HCT116 colon tumor xenografts. Human HCT116 tumors were created by s.c. injection of cells in Harlan athymic mice. (A and B) Tumors were studied by FLT-PET as described in the Materials and Methods section, immediately before (day 0) and on days 3 and 10 after treatment with vehicle or everolimus (10 mg/kg p.o.). Results show (n = 6 per treatment group) the individual SUVs for tumors (A) and the mean ± SEM fractional effect of treatment (B). (C) Histology and IHC of ablated tumors on day 10 after completion of the second PET scan.

Article Snippet: Female Brown-Norway rats and C57BL/6 mice were obtained from Charles River (France) and female Harlan Hsd:Npa nu/nu (nude) athymic mice were obtained from the Novartis breeding stock (Basel, Switzerland).

Techniques: Injection

Journal: Genes & Development

Article Title: Prolyl hydroxylation by EglN2 destabilizes FOXO3a by blocking its interaction with the USP9x deubiquitinase

doi: 10.1101/gad.242131.114

Figure Lengend Snippet: Differential binding of USP9x underlies control of FOXO3a stability by EglN2. ( A ) Immunoblot analysis of T47D cells that were infected with a lentivirus encoding the indicated USP9x shRNAs or Ctrl shRNA. ( B ) Immunoblot analysis of T47D cells that were infected with a lentivirus encoding HA-FOXO3a and then superinfected with a lentivirus encoding USP9x shRNA (364 or 064) or Ctrl shRNA. ( C ) Immunoblot analysis of 293T cells that were infected with a lentivirus encoding USP9x shRNA (364) or Ctrl shRNA and then, after drug selection, transfected with a plasmid encoding V5-tagged mouse full-length USP9x or empty vector (Ctrl). ( D ) Immunoblot (IB) assays of whole-cell extracts (WCE) and immunoprecipitates (IP) of T47D cells that were infected with a lentivirus encoding USP9x shRNA (sh128 or sh364) or Ctrl shRNA and then, after drug selection, transiently transfected with plasmids encoding Flag-ubiquitin and HA-FOXO3a followed by treatment with 10 μM MG132 for 16 h. ( E ) Immunoblot analysis of MCF-7 cells that were infected with a lentivirus encoding shRNA against either USP9x (364) or Ctrl shRNA followed by 100 μg/mL cycloheximide for the indicated duration. ( F ) Immunoblot analysis of 293T cells that were cotransfected with plasmids encoding V5-tagged USP9x (wild-type or C1556S) and HA-FOXO3a. ( G ) Immunoblot (IB) assays of whole-cell extracts (WCE) and immunoprecipitates (IP) of T47D cells that were transfected with a plasmid encoding HA-FOXO3a (wild-type or P426A;P437A) followed by treatment with 10 μM MG132 for 16 h. ( H ) Immunoblot analysis of bound USP9x recovered from lysed T47D cells that were incubated with 20 μL of Neutravidin-Sepharose preloaded with the indicated FOXO3a peptides. ( I , J ) Immunoblot analysis of T47D cells that were infected with a lentivirus encoding USP9x shRNA (364) or Ctrl shRNA and then, after drug selection, treated with the indicated drugs or siRNAs.

Article Snippet: V5-tagged mouse USP9x, catalytic-dead USP9x C1556S, and GST-tagged human USP9x (N1, N2, C1, and C2) plasmids were generously provided by Dr. Feng Cong (Novartis).

Techniques: Binding Assay, Western Blot, Infection, shRNA, Selection, Transfection, Plasmid Preparation, Incubation

Journal: Genes & Development

Article Title: Prolyl hydroxylation by EglN2 destabilizes FOXO3a by blocking its interaction with the USP9x deubiquitinase

doi: 10.1101/gad.242131.114

Figure Lengend Snippet: Proposed models for FOXO3a regulation by EglN2. ( A ) Estrogen binding with ER transcriptionally induces EglN2, which triggers FOXO3a hydroxylation on prolyl residue 426 and 327 sites. Hydroxylation of these sites dissociates FOXO3a from USP9x deubiquitinase, thereby facilitating FOXO3a ubiquitylation and degradation. ( B ) Depletion of EglN2 by either lack of ER activity or EglN2 siRNAs/shRNAs prevents FOXO3a from being hydroxylated. As a result, USP9x binds with FOXO3a and prevents FOXO3a from being ubiquitylated and degraded. Stabilization of FOXO3a decreases Cyclin D1 expression.

Article Snippet: V5-tagged mouse USP9x, catalytic-dead USP9x C1556S, and GST-tagged human USP9x (N1, N2, C1, and C2) plasmids were generously provided by Dr. Feng Cong (Novartis).

Techniques: Binding Assay, Activity Assay, Expressing