Journal: ASN NEURO

Article Title: Diazepam Binding Inhibitor Control of Eu- and Hypoglycemic Patterns of Ventromedial Hypothalamic Nucleus Glucose-Regulatory Signaling

doi: 10.1177/17590914231214116

Figure Lengend Snippet: Effects of Dbi Gene Silencing on Eu- or Hypoglycemic Neurotransmitter and Metabolic Sensor Marker Gene Expression Patterns in VMNvl Nitrergic Neurons. Groups (n = 4/group) of Dbi or SCR siRNA-pretreated, V- or INS-injected male rats were sacrificed one hour after sc injection. Individual VMNdm neuronal nitric oxide synthase (nNOS/NOS1)-immunopositive neurons were laser-catapult-microdissected from 10 micron-thick fresh frozen sections for multiplex single-cell qPCR analyses. Data depict mean normalized nNOS/NOS1 , Gck , Gkrp , AMPKα1/Prkaa1 , or AMPKα2/Prkaa2 mRNA measures ± S.E.M. for SCR siRNA/V (n = 8), Dbi siRNA/V (n = 8), SCR siRNA/INS (n = 8), Dbi siRNA/INS (n = 8) treatment groups. Outcomes of statistical analyses are as follows: nNOS: F (3,28) : 365.90, p < 0.001; Knockdown main effect: F (1,28) : 337.27, p < 0.0001; INS main effect: F (1,28) : 483.78; p < 0.001; Knockdown/INS interaction: F (1,28) : 276.65; p < 0.001; Gck: F (3,28) : 328.81, p < 0.001; Knockdown main effect: F (1,28) : 822.64, p < 0.0001; INS main effect: F (1,28) : 7.36, p = 0.011; Knockdown/INS interaction: F (1,28) : 156.42, p < 0.001; Gkrp: F (3,28) : 93.08, p < 0.001; Knockdown main effect: F (1,28) : 173.19, p < 0.001; INS main effect: F (1,28) : 92.90, p < 0.001; Knockdown/INS interaction: F (1,28) : 13.14, p = 0.001; AMPKα1/Prkaa1: F (3,28) : 122.73, p < 0.001; Knockdown main effect: F (1,28) : 24.09, p < 0.0001; INS main effect: F (1,28) : 218.50, p < 0.001; Knockdown/INS interaction: F (1,28) : 125.60, p < 0.001; AMPKα2/Prkaa2: F (3,28) : 160.07, p < 0.001, Knockdown main effect: F (1,28) : 168.68, p < 0.0001; INS main effect: F (1,28) : 121.08, p < 0.001; Knockdown/INS interaction: F (1,28) : 190.44, p < 0.001. Statistical differences between discrete pairs of treatment groups are denoted as follows: * p < 0.05; ** p < 0.01; and *** p < 0.001.

Article Snippet: Briefly, after ice-cold acetone fixation (5 min), sections were blocked (1 h) with 1.5% normal goat serum (prod. no. S-1000, Vector Laboratories, Burlingame, CA, U.S.A.) in Tris-buffered saline, pH 7.4, (TBS) containing 0.05% Triton X-100 prior to incubation (36–48 h, 4°C) with rabbit anti-rat Gad 65/67 (Gad; prod. no. ABN904, 1:1000; Millipore Sigma, Burlington, MA, U.S.A.; RRID:AB_2893025) or anti-rat nNOS (prod. no. NBP1-39681, 1:1000; Novus Biologicals, Littleton, CO, U.S.A.; RRID:AB_2282822) primary antibody ( ; ; ; ).

Techniques: Marker, Gene Expression, Injection, Multiplex Assay, Knockdown

Journal: ASN NEURO

Article Title: Diazepam Binding Inhibitor Control of Eu- and Hypoglycemic Patterns of Ventromedial Hypothalamic Nucleus Glucose-Regulatory Signaling

doi: 10.1177/17590914231214116

Figure Lengend Snippet: Eu- versus Hypoglycemic Neurotransmitter and Metabolic Sensor Marker Gene Expression Patterns in VMNdm Nitrergic Neurons: Effects of Dbi Gene Knockdown. Experimental design, treatment group designations, target gene profiles, analytical and statistical methods, and data presentation format are identical to that described in above, except that nNOS-immunopositive neurons were laser-catapult-microdissected from the VMNdm. Outcomes of statistical analyses are as follows: nNOS: F (3,28) : 78.52, p < 0.001; Knockdown main effect: F (1,28) : 65.83, p < 0.0001; INS main effect: F (1,28) : 526.22, p < 0,001; Knockdown/INS interaction: F (1,28) : 113.52, p < 0.001; Gck: F (3,28) : 859.64, p < 0.001; Knockdown main effect: F (1,28) : 531.39, p < 0.0001; INS main effect: F (1,28) : 1197.25, p < 0.001; Knockdown/INS interaction: F (1,28) : 850.27, p < 0.001; Gkrp: F (3,28) : 14.82, p < 0.001; Knockdown main effect: F (1,28) : 1.04, p = 0.317; INS main effect: F (1,28) :4.44, p = 0.044; Knockdown/INS interaction: F (1,28) : 38.99, p < 0.001; AMPKα1/Prkaa1: F (3,28) : 39.14, p < 0.001; Knockdown main effect: F (1,28) : 14.03, p = 0.001; INS main effect: F (1,28) : 20.78, p < 0,001; Knockdown/INS interaction: F (1,28) : 82.60, p < 0.001; AMPKα2/Prkaa2: F (3,28) : 88.86, p < 0.001; Knockdown main effect: F (1,28) : 46.27, p < 0.0001; INS main effect: F (1,28) : 183.48, p < 0.001; Knockdown/INS interaction: F (1,28) : 36.81, p < 0.001. *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: Briefly, after ice-cold acetone fixation (5 min), sections were blocked (1 h) with 1.5% normal goat serum (prod. no. S-1000, Vector Laboratories, Burlingame, CA, U.S.A.) in Tris-buffered saline, pH 7.4, (TBS) containing 0.05% Triton X-100 prior to incubation (36–48 h, 4°C) with rabbit anti-rat Gad 65/67 (Gad; prod. no. ABN904, 1:1000; Millipore Sigma, Burlington, MA, U.S.A.; RRID:AB_2893025) or anti-rat nNOS (prod. no. NBP1-39681, 1:1000; Novus Biologicals, Littleton, CO, U.S.A.; RRID:AB_2282822) primary antibody ( ; ; ; ).

Techniques: Marker, Gene Expression, Knockdown

Journal: British Journal of Pharmacology

Article Title: Serelaxin-mediated signal transduction in human vascular cells: bell-shaped concentration–response curves reflect differential coupling to G proteins

doi: 10.1111/bph.12964

Figure Lengend Snippet:

Article Snippet: Briefly, 20–30 μg of protein was separated on a polyacrylamide gel, transferred to a PVDF membrane and probed with the following primary antibodies: anti-eNOS antibody (R&D Systems; 1:1000, overnight at 4°C), anti-nNOS antibody (R&D Systems; 1:500, overnight at 4°C), anti-iNOS antibody (Cell Signalling, Beverley, MA, USA; 1:1000, overnight at 4°C), anti-VEGF antibody (Abcam, Cambridge, MA, USA; 1:1000, 2 h at room temperature), anti-ET B receptor antibody (Pierce, Rockford, IL, USA; 1:1000, overnight at 4°C), anti-β-actin antibody (Cell Signalling; 1:1000, 2 h at room temperature).

Techniques: