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Dating laurie palanza

ionophore (100 ng/ml, ionomycin), soluble anti-CD3ɛ (0.1–1 μg/ml, clone 145–2C11), anti-CD28 (0.02–0.2 μg/ml, clone 37.51), or recombinant mouse IL-2 (50 units/ml) in triplicate for 12, 24, 48, 72, or 120 h, followed by a 12-h pulse with [ cells) by negative magnetic sorting and activated with various stimuli.

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Single-cell suspensions of thymi, lymph nodes, and spleens were stained with FITC-, phycoerythrin-, or biotin-conjugated Abs reactive to CD3ɛ, TCRαβ, CD4, CD8, CD25, CD44, CD28, CD69, CD11b (Mac-1), CD11c, B220, CD43, s Ig M, s Ig D, or Gr-1.Several objects were placed as cues around the maze.On day 2, each subject was again placed in the T-maze and given access to the left arm of the maze.Four- to five-month-old sex-matched littermate mice were examined for neuromotor functions (22): vibrissae, eyes, rearing/support/standing, muscle tone, tail hanging-induced turns and limbs/digits extension, ear reflex, eye blink and pupillary response to light, sweet/bitter tasting acceptance/rejection, and orientation toward sound.Circadian and open-field activities (i.e., cage crosses, total distance, vertical movements) were measured in cages fitted with infrared sensors. Motor coordination was analyzed by a rotating rod task.Time to find the platform and total distance were measured.

In the T-maze test, on day 1 the mice were placed in an empty maze for 10 min with access only to its central and right arms.

Recently it has been shown that RGS9–1 controls the photosensitization response in the mouse eye (11).

Axin, which contains a RGS domain, negatively regulates the Wnt-signaling pathway, and -mutant mice display defects in embryonic axis formation (12).

Targeted ES cell clones were injected into C57BL/6 blastocysts and heterozygous mice obtained by mating chimeras with C57BL/6J mice.

PCR primers: wild-type allele (5′-CCGAGTTCTGTGAAGAAAACATTG3′; 5′GGGACTCCTGGTCTCATGTAGCAT3′) and mutant allele (5′-GCTAAAGCGCATGCTCCAGAC-3′; 5′-GGCCCACATTTACACGAACC-3′).

The first RGS identified was the yeast protein Sst2, isolated in a screen for mutants that failed to down-regulate the GPCR-mediated response to mating pheromone (8–9).