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Figure 5 shows the corresponding load displacement (L D) curves of the AAO specimen at indentation loads of 1, 3 and 5 mN at room temperature.

The sections were then washed in PBS for 30 mn and incubated with Streptavidin peroxydase (Dako) diluted 1/100 in PBS, BSA 1% for 45 mn at room temperature.

AP was detected using NBT and BCiP for 30 mn at room temperature.

The slides were then rinsed and fixed by incubating in 4% paraformaldehyde for 20 mn at room temperature.

Cryostat sections were rinsed twice for 5 mn at room temperature in Neurobasal medium (Gibco) containing 0.09% glucose, 0.2% Bovine Serum Albumin (BSA) and 0.02% bacitracin.

DNA, in a buffer solution, was passed through a 5.0 μm Cameo 30N filter (Osmonics, Minnetonka, MN) at room temperature before precipitation and then washed with ice-cold 70% ethanol.

Antibody detection was with FITC-conjugated goat anti-mouse IgG (7.5 μg ml−1, Jackson Laboratories, Bar Harbor, MN, USA) for 45 min at room temperature.

The slides were then incubated with a mouse monoclonal anti-human FXR antibody (1 30 dilution; R&D Systems Co., Minneapolis, MN, USA) for 90 min at room temperature.

Cells were stained with a FITC-labeled anti-DR4 antibody (10  μg/ml, Axxora, Loerrach, Germany), a FITC-labeled anti-DR5 antibody (10  μg/ml; Axxora) or a FITC-labeled IgG1-isotype control antibody (10  μg/ml; Ancell, Bayport, MN, USA) for 60 min at room temperature.

MgxMn(1−x)(BH4)2 (x = 0 0.8) conserves the trigonal structure of Mn(BH4 2 at room temperature.

Samples (n = 3) were equilibrated for 1 min under 1 mN load at room temperature.