Journal: PLoS ONE

Article Title: Allopregnanolone Preclinical Acute Pharmacokinetic and Pharmacodynamic Studies to Predict Tolerability and Efficacy for Alzheimer’s Disease

doi: 10.1371/journal.pone.0128313

Figure Lengend Snippet: A. In 5-month-old 3xTgAD mouse hippocampus, BrdU+ nuclei increased significantly in 24 h after a single subcutaneous (SC) dose of Allo 0.5, 1, or 10 mg/kg. B. In 5-month-old 3xTgAD mouse hippocampus, protein expression of ~29 kDa PCNA increased significantly at 24 h after SC Allo 0.5 and 1 mg/kg doses whereas 10 mg/kg dose trended towards increase but did not reach significance. Transdermal and subcutaneous Allo increased PCNA protein expression in male mouse AD model. C. In 5-month-old 3xTgAD mouse hippocampus, protein expression of PCNA increased significantly at 4 h after transdermal (TD) Allo 50 mg/kg and SC Allo 10 mg/kg doses. D. In 15-month-old nonTg mouse hippocampus, protein expression of PCNA increased significantly at 4 h after TD Allo 50 mg/kg dose. E. In 5-month-old 3xTAD and 15-month-old nonTg mouse hippocampus, BrdU+ nuclei increased significantly in 24 h after intranasal (IN) dose of Allo 3 mg/kg 100% Castor Oil and Allo 10 mg/kg 20% HBCD suspension doses. F. In 5-month-old 3xTgAD and 15-month-old nonTg mouse hippocampus, protein expression of PCNA increased significantly at 24 h after IN Allo 3 mg/kg 100% Castor Oil and Allo 10 mg/kg 20% HBCD suspension doses. Intramuscular Allo-induced increase in cell cycle marker in male mouse AD model. G. In 5-month-old 3xTgAD mouse hippocampus, BrdU+ nuclei increased significantly in 24 h after a single intramuscular (IM) dose of Allo 2 mg/kg and SC Allo 10 mg/kg dose. H. In 5-month-old 3xTgAD mouse hippocampus, protein expression of PCNA increased significantly at 24 h post-IM Allo 2 mg/kg dose and SC Allo 10 mg/kg dose. Intravenous Allo-induced increase in cell cycle and neurodifferentiation markers in male mouse AD model. I. In 5-month-old 3xTgAD mouse hippocampus, protein expression of 30 kDa cyclinD2 increased significantly at 4 h post-intravenous (IV) Allo 0.1 and 0.5 mg/kg dose whereas 1 mg/kg dose trended towards increase but did not reach significance. J. In 5-month-old 3xTgAD mouse hippocampus, protein expression of PCNA increased significantly at 4 h after IV Allo 0.5 mg/kg dose whereas 0.1 and 1 mg/kg dose did not reach significance. K. In 5-month-old 3xTgAD mouse hippocampus, protein expression of ~40 kDa doublecortin (DCX) increased significantly at 4 h after IV Allo 0.5 and 1 mg/kg doses. L. In 5-month-old 3xTgAD mouse hippocampus, protein expression of 49 kDa NeuroD increased significantly at 4 h after IV Allo 0.5 mg/kg, whereas 0.1 and 1 mg doses did not reach significance. Intravenous Allo-induced rapid transient increase in CREB phosphorylation in male mouse aging model. M. In 15-month-old nonTg mouse hippocampus, protein expression of 43 kDa serine 133 phosphorylated CREB (pCREB) increased significantly 5 min after IV Allo 1.5 mg/kg dose. N. In 15-month-old nonTg mouse hippocampus, protein expression of 49 kDa NeuroD1 (NeuroD) increased significantly at 4 h then decreased at 24 h after intravenous Allo 1.5 mg/kg dose. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001, n = 4–6, bars represent mean value ± SEM.

Article Snippet: The following antibodies were used in this study: NeuroD (1:1000, Cell Signaling Technology), Doublecortin (1:300, AbCam), proliferating cell nuclear antigen (PCNA; 1:1000, Millipore), CyclinD2 (1:1000, Cell Signaling Technology), pCREB (1:1000, Cell Signaling Technology), CREB (1:1000, Cell Signaling Technology).

Techniques: Expressing, Marker

Journal: PLoS ONE

Article Title: Allopregnanolone Preclinical Acute Pharmacokinetic and Pharmacodynamic Studies to Predict Tolerability and Efficacy for Alzheimer’s Disease

doi: 10.1371/journal.pone.0128313

Figure Lengend Snippet: Summary of mouse efficacy data and statistical comparisons.

Article Snippet: The following antibodies were used in this study: NeuroD (1:1000, Cell Signaling Technology), Doublecortin (1:300, AbCam), proliferating cell nuclear antigen (PCNA; 1:1000, Millipore), CyclinD2 (1:1000, Cell Signaling Technology), pCREB (1:1000, Cell Signaling Technology), CREB (1:1000, Cell Signaling Technology).

Techniques: Biomarker Assay

Journal: British Journal of Cancer

Article Title: CDK4/6 inhibition presents as a therapeutic option for paediatric and adult germ cell tumours and induces cell cycle arrest and apoptosis via canonical and non-canonical mechanisms

doi: 10.1038/s41416-020-0891-x

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: CyclinD2 , BD Pharmingen , G132-43 , 554200 , 1:500 , Western blot.

Techniques: Western Blot, Immunohistochemistry, Immunofluorescence, Flow Cytometry

Journal: The Journal of Biological Chemistry

Article Title: Forkhead Box Protein 1 (FoxO1) Inhibits Accelerated β Cell Aging in Pancreas-specific SMAD7 Mutant Mice *

doi: 10.1074/jbc.M116.770032

Figure Lengend Snippet: β Cell dysfunction and aging in SMAD7Ptf1a mice. A–G, body weight (A), fasting blood glucose (B), IPGTT (C and D), serum insulin (E), β cell mass (F), and serum glucagon (G) at different ages in SMAD7Ptf1a mice compared with littermate control SMADfx/fx mice. H, 1 week of BrdU labeling followed by quantification of the percentage of β cells that incorporated BrdU. I and J, enrichment of gene transcripts of CyclinD1 (I) and CyclinD2 (J) in mouse islets of various ages. The values were normalized against CycloA, which is consistent among all conditions. K, islet perifusion study using islets from 20-week SMAD7Ptf1a and SMAD7fx/fx mice. Impaired glucose-stimulated insulin secretion was detected in SMAD7Ptf1a mice. *, p < 0.05 and n = 5 in all cases.

Article Snippet: Primary antibodies were as follows: guinea pig polyclonal insulin-specific (Dako, Carpinteria, CA); goat polyclonal NeuroD1-specific (Santa Cruz Biotechnology, Dallas, TX); rabbit polyclonal SMAD7-, CyclinD1-, and CyclinD2-specific (Santa Cruz Biotechnology); FoxO1- and GAPDH-specific (Cell Signaling Technology, San Jose, CA), Collagen I- and Pdx1-specific (Abcam, Cambridge, MA, USA), MafA-specific (Bethyl Laboratories, Inc.); Nkx6.1-specific (a kind gift from Dr. Maike Sander, University of California, San Diego, CA); and rat CD31-specific (BD Biosciences) and BrdU-specific (Abcam).

Techniques: Control, Labeling