On day 7, a final probe trial (60 s in the pool without platform) was applied.
However, Axin has no detectable GAP activity for Gα (10).
The physiological roles of other mammalian RGS proteins are yet to be established.
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.
Basal serum Ig levels (Ig M, Ig G1, Ig G2a, Ig G2b, Ig G3, and Ig A) were determined by ELISA (20).
For lymphocytic choriomeningitis virus (LCMV) infections, mice were infected with LCMV (700 plaque-forming units, s.c.) into the hind footpad.
Cellular infiltration was measured daily as described (21).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′).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.mice also display increased anxiety responses and decreased male aggression in the absence of cognitive or motor deficits.