Journal: The Journal of Neuroscience

Article Title: Suppression of Postischemic Hippocampal Nerve Growth Factor Expression by a c- fos Antisense Oligodeoxynucleotide

doi: 10.1523/JNEUROSCI.19-04-01335.1999

Figure Lengend Snippet: The presence of bio-s-ODN in the rat brain. Bio-s-ODN (1 nmol) with lipofectin (0.01 μg) was delivered via intracerebroventricular route as described previously (Liu et al., 1994). The presence of bio-s-ODN in the brain was detected by using goat IgG against biotin and rabbit anti-goat IgG-FITC conjugates. The nuclei were counterstained with PI and appeared red. Representative areas in layer II of the cortex (A,B, and C at 6, 48, and 96 hr after delivery, respectively) and in CA1 of the hippocampus (D, E, and F at 6, 24, and 96 hr after delivery, respectively) in the right hemisphere are shown. The magnification in all panels is the same. Scale bar, 20 μm. The image was acquired using double exposure and Kodak photographic film for FITC and PI signals. Because of the double staining of FITC and PI, the biotin signal appearedgreen (F) or yellow(a blend of red and green;B, C, E). The apparent larger sizes of the nuclei in B–F were attributable to the visualization of bio-s-ODNs in cell bodies in contrast to nuclear stain only by PI in A.

Article Snippet: The presence of s-ODNs was detected using 3′-OH-terminus labeling of TdT-mediated dig-dUTP incorporation, followed by immunohistochemical staining using anti-dig-IgG-FITC conjugates (Oncor, Inc.).

Techniques: Double Staining, Staining

Journal: The Journal of Neuroscience

Article Title: Suppression of Postischemic Hippocampal Nerve Growth Factor Expression by a c- fos Antisense Oligodeoxynucleotide

doi: 10.1523/JNEUROSCI.19-04-01335.1999

Figure Lengend Snippet: The uptake of s-ODN in the ependyma of the lateral ventricle and beyond. The presence of s-ODNs was detected using 3′-OH-terminus labeling of TdT-mediated dig-dUTP incorporation, followed by immunohistochemical staining using anti-dig-IgG-FITC conjugates (Oncor, Inc.). The images were acquired using the SPOT digital camera with a single exposure using the green (FITC) spectrum. The anti-dig-FITC–dig-dUTP complex is shown as whitefluorescence. Four different doses of c-fos antisense s-ODNs, each administered with 10 μg of lipofectin (1 of the 3 animals from each group is shown) in 40 μl of aCSF (A, saline with no s-ODN; B, 1 nmol; C, 10 nmol). One of the three animals from each dosage is shown. The uptake time was 4 hr after infusion.

Article Snippet: The presence of s-ODNs was detected using 3′-OH-terminus labeling of TdT-mediated dig-dUTP incorporation, followed by immunohistochemical staining using anti-dig-IgG-FITC conjugates (Oncor, Inc.).

Techniques: Labeling, Immunohistochemical staining, Staining

Journal: The Journal of Neuroscience

Article Title: Suppression of Postischemic Hippocampal Nerve Growth Factor Expression by a c- fos Antisense Oligodeoxynucleotide

doi: 10.1523/JNEUROSCI.19-04-01335.1999

Figure Lengend Snippet: Representative digitized images of brain slices showing the uptake of s-ODN. The presence of s-ODNs in the dentate gyrus was detected using 3′ end labeling as in Figure ​Figure2,2, except that 10 nmol of c-fos antisense s-ODN with 10 μg of lipofectin in 40 μl of aCSF was delivered, and the images were acquired as in Figure ​Figure2.2. The animals were killed at 16 hr after infusion. The anti-dig-FITC–dig-dUTP complex is shown as bright whitefluorescence. Dark dots appear in the dentate gyrus of the animals that received s-ODN but without TdT for dUTP incorporation during the 3′-OH end labeling assay (A). Scale bar, 20 μm.

Article Snippet: The presence of s-ODNs was detected using 3′-OH-terminus labeling of TdT-mediated dig-dUTP incorporation, followed by immunohistochemical staining using anti-dig-IgG-FITC conjugates (Oncor, Inc.).

Techniques: End Labeling