Journal: Experimental Biology and Medicine

Article Title: Liraglutide prevents fast weight gain and β-cell dysfunction in male catch-up growth rats

doi: 10.1177/1535370214567614

Figure Lengend Snippet: Effects of HFD catch-up growth and liraglutide treatment on β-cell function and apoptosis. Pdx-1 protein (a1) and mRNA (a2) levels of islets at the end of week 8. Pdx-1 protein levels of islets were assessed by Western blot and the results were expressed as the ratios of Pdx-1 and GAPDH. The Pdx-1 mRNA levels were detected by qPCR and expressed as the ratios of Pdx-1 and β-actin. Protein levels of Bcl-2 (b1) and Caspase-3 p 12 subunit (c1) were detected by Western blot and the results were expressed as the ratios of Caspase-3 p12 subunit or Bcl-2 with GAPDH. The mRNA levels of Bcl-2 (b2) and Procaspase-3 (c2) were expressed as the ratios of Proaspase-3 or Bcl-2 with β-actin. All the results are expressed as mean ± SEM. *p < 0.01 versus the NC group; #p < 0.01 versus the CUG group; &p < 0.01 versus the CUGL group

Article Snippet: Primer sequences Western blot analysis Protein extracts from islets (20 μg from each rat) were subjected to Western blot analysis using mouse antipancreatic duodenal homeobox-1 (Pdx-1) monoclonal antibody (1:800; Cell Signaling Technology, Danvers, MA, USA), mouse anti-Caspase-3 p12 subunit monoclonal antibody (1:500, Boster Bio-Engineering, China), mouse anti-B-cell lymphoma (Bcl-2) polyclonal antibody (1:800, Bioworld, USA), and mouse anti-GAPDH antibody (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Cell Function Assay, Western Blot

Journal: bioRxiv

Article Title: Obesity alters mobility and adult neurogenesis, but not hippocampal dependent learning in ob/ob mice

doi: 10.1101/537720

Figure Lengend Snippet: Adult neurogenesis, proliferation and apoptosis were analyzed using immunofluorescence staining within the dentate gyrus (DG) of the hippocampus. Cell numbers of wild type (wt) animals were set to 100 %. A The number of proliferating cells was visualized using phosphohistone H3 (PH3). Leptin-deficient (ob/ob) mice showed a reduced amount of PH3-positive proliferating cells. B As a maker for immature neuronal cells, doublecortin (DCX) was used. Ob/ob mice show a reduced number of DCX positive cells in the DG compared to wild type controls. C As an apoptosis marker caspase3 (Casp3) was used. The number of Casp3 positive cells did not differ between the two groups of mice. Fluorescence images were taken using a Olympus BX63 microscope with 25,2x magnification (40x objective, 0,63x camera) in the DG of wt mice for D PH3 positive, E DCX positive and F Casp3 positive cells.

Article Snippet: After washing with PBS, the solution containing the primary rabbit anti-active Caspase 3 antibody (Merck Millipore, Germany) was applied 1:100 in blocking serum (3 % NGS + 0.1 % Triton X-100 in PBS) and incubated at 4°C over night.

Techniques: Immunofluorescence, Staining, Marker, Fluorescence, Microscopy

Journal: bioRxiv

Article Title: Evolution of lbx spinal cord expression and function

doi: 10.1101/2020.12.30.424885

Figure Lengend Snippet: (a-c) Lateral views of activated caspase-3 immunohistochemistry in zebrafish spinal cord ( a & b ) or whole embryo ( c ) at 30h in (a) WT embryo, (b) lbx1a sa1496 ;lbx1b hu3534 double mutant and (c) smoothened b641 mutant. The latter was used as a positive control as apoptosis is increased in the head and tail regions. In all cases anterior is left, dorsal top, (a & b) White arrow heads indicate Caspase-3-positive cells. (d) Numbers of Caspase-3-positive cells in spinal cord region adjacent to somites 6-10 in lbx1a sa1496 ;lbx1b hu3534 double mutants and WT siblings. Values shown are the mean from 3 different embryos, the S.E.M. and the P value from a student’s t-test. Scale bar = 50 μm (a & b) and 200 μm (c) .

Article Snippet: Primary antibodies used were chicken polyclonal anti-GFP (Abcam, ab13970, 1:500), rabbit anti-GFP (Molecular Probes A6465, 1:500) and rabbit anti-activated Caspase-3 (Fisher Scientific/BD, BDB559565, 1:500).

Techniques: Immunohistochemistry, Mutagenesis, Positive Control