Journal: World Journal of Gastroenterology

Article Title: Translocator protein facilitates neutrophil-mediated mucosal inflammation in inflammatory bowel diseases

doi: 10.3748/wjg.v31.i27.109239

Figure Lengend Snippet: Translocator protein expression is significantly upregulated in mice with colitis, n = 6 in each group. A: The body weight of mice receiving H 2 O or dextran sodium sulfate (DSS) is monitored weekly. Changes in body weight are expressed as a percentage of the original weight at the beginning of the experiments. a P < 0.05, b P < 0.01 vs mice those received H 2 O at the end of the 8 th week; B: Changes in disease activity index score between the two groups during the experiment; C: Gross morphology of the large bowels at the end of the 8 th week after receiving DSS; D: Colon length is measured and recorded. b P < 0.01 vs mice those received H 2 O; E: Histological appearance of colonic sections after hematoxylin and eosin. Original magnification 200 ×; F: Pathological scores of colonic sections are shown in the chart; G: Representative immunofluorescent images of the colonic mucosa for staining with 4'-6-diamidino-2-phenylindole (blue), anti-TSPO (red), and anti-myeloperoxidase (MPO) (green). The white arrows indicate representative TSPO + MPO + cells. Original magnification 400 ×; H: Representative [ 18 F] labeled N,N-diethyl-2-(2-[4-(2-fluoroethoxy)phenyl]-5,7-dimethylpyrazolo[1,5-alpha]pyrimidine-3-yl)acetamide (DPA-714) uptake according to micro-positron emission tomography-computed tomography imaging in wild-type (WT) mice with H 2 O and those with DSS treatment. Colon [ 18 F]DPA-714 uptake is higher in the WT mice with DSS treatment than in those with H 2 O treatment. Regions of interest drawn in the colon tissue are showed as white arrows on each image. n : The biologically independent replicates used in the statistical analyses. DSS: Dextran sodium sulfate; TSPO: Translocator protein; MPO: Myeloperoxidase; DAPI: 4'-6-diamidino-2-phenylindole.

Article Snippet: Coverslips were mounted using the Slowfade Gold antifade mountant with DAPI (Servicebio, 1:1000) for DNA counterstaining.

Techniques: Expressing, Activity Assay, Staining, Labeling, Micro-PET, Computed Tomography, Imaging

Journal: World Journal of Gastroenterology

Article Title: Translocator protein facilitates neutrophil-mediated mucosal inflammation in inflammatory bowel diseases

doi: 10.3748/wjg.v31.i27.109239

Figure Lengend Snippet: Dextran sodium sulfate-induced colitis leads to changes in translocator protein expression and several types of neural cells in the brain. A and B: Representative immunofluorescent images for staining with 4'-6-diamidino-2-phenylindole (DAPI) (blue), anti-translocator protein (TSPO) (red), and anti-glial fibrillary acidic protein (green) in the hippocampus and cerebral cortex of representative sections from the brain of two groups. Original magnification: (1) Up 200 ×; and (2) Down 400 ×; C and D: Representative immunofluorescent images for staining with DAPI (blue), anti-TSPO (red), and anti-IBA1 (green) in the hippocampus and cerebral cortex of representative sections from the brain of two groups. Original magnification: (1) Up 200 ×; and (2) Down 400 ×. n : The biologically independent replicates used in the statistical analyses. DAPI: 4'-6-diamidino-2-phenylindole; DSS: Dextran sodium sulfate; GFAP: Glial fibrillary acidic protein; TSPO: Translocator protein.

Article Snippet: Coverslips were mounted using the Slowfade Gold antifade mountant with DAPI (Servicebio, 1:1000) for DNA counterstaining.

Techniques: Expressing, Staining

Journal:

Article Title: Guanosine supplementation reduces apoptosis and protects renal function in the setting of ischemic injury

doi:

Figure Lengend Snippet: Effects of guanosine and enhanced GTP recovery on renal cell apoptosis at 24 hours. Mice were treated as detailed in legend of Figure ​Figure2.2. Kidney sections were obtained at 24 hours and costained with TUNEL and DAPI. (a) A negative control (NC; TdT omitted) and (b) a DNase-treated positive control (PC) for TUNEL. All TUNEL-positive nuclei had bright green fluorescence. (c) A representative field showing the medulla from NS/I group. Extensive TUNEL-positive staining is seen and is localized primarily to the tubular epithelium (dark green autofluorescence). (d) A representative field from the medulla of the G/I group showing lack of TUNEL-positive staining. (e) A magnification of TUNEL-positive nuclei from NS/I mice, with only the DAPI channel turned on. ×60. Nuclei are condensed, dysmorphic, and show heavy staining. Inset shows a single nucleus. ×120. Typical apoptotic fragmentation of chromatin into four small bodies is seen. (f) A magnification of TUNEL-negative nuclei from G/I mice. ×60. They show lack of condensation and faint DAPI staining.

Article Snippet: Finally, separate sections were stained with TUNEL reagent (Promega Corp., Madison, Wisconsin, USA) and DAPI for in situ apoptosis detection.

Techniques: TUNEL Assay, Negative Control, Positive Control, Fluorescence, Staining

Journal:

Article Title: Guanosine supplementation reduces apoptosis and protects renal function in the setting of ischemic injury

doi:

Figure Lengend Snippet: Quantitative evaluation of TUNEL-positive nuclei in cortex and medulla. Values are means ± SE and represent number of TUNEL-positive nuclei expressed as percentage of total number of nuclei. Mice were treated as detailed in the legend of Figure ​Figure2.2. Kidney sections were obtained at 24 hours and processed for TUNEL and DAPI staining as detailed in Methods. Number of fields counted: 20 for G/I, 35 for NS/I, 8 for G/sham, and 8 for NS/sham. (*P < 0.01 when medullary TUNEL-positive nuclei in NS/I were compared to all other groups).

Article Snippet: Finally, separate sections were stained with TUNEL reagent (Promega Corp., Madison, Wisconsin, USA) and DAPI for in situ apoptosis detection.

Techniques: TUNEL Assay, Staining

Journal:

Article Title: Guanosine supplementation reduces apoptosis and protects renal function in the setting of ischemic injury

doi:

Figure Lengend Snippet: Distribution of TUNEL-positive nuclei among various nephronal segments. (a) A 5-μm thick section from NS/I mice stained with TUNEL and DAPI. The TUNEL positive nuclei were pseudocolored white instead of green to allow clear distinction from the green FITC-phalloidin. (b) A 5-μm section immediately contiguous to the one shown in a is stained with the actin marker FITC-phalloidin (bright green) to identify proximal tubules (P). Distal tubules (D) show minimal or no staining. The section was costained with anti-THP Ab and a Cy5-conjugated secondary Ab (red). This is predominantly a marker for Henle’s loop segments (LH). (c) An overlay of a and b allows the localization of TUNEL-positive nuclei to various tubular segments. (d) A quantitative estimate of the distribution of TUNEL-positive nuclei normalized to total number of DAPI-positive nuclei for each particular tubular segment or lumen. Other refers to interstitial or vascular TUNEL-positive nuclei.

Article Snippet: Finally, separate sections were stained with TUNEL reagent (Promega Corp., Madison, Wisconsin, USA) and DAPI for in situ apoptosis detection.

Techniques: TUNEL Assay, Staining, Marker