For adipogenic induction, the cells were washed twice with PBS (Phosphate buffered saline) and treated in a defined Adipogenic Induction Medium [AIM: DMEM-LG supplemented with 10% FBS, 50 µg/ml ascorbate-2 phosphate (Sigma, St . Louis MO), 10−7 M dexamethasone (Sigma), 50 µg/ml indomethacin (Sigma), and 10 µg/ml insulin (Sigma)].
In brief, after rehydrated in water, the paraffin sections placed in citric buffer (pH 6.0) and treated in a microwave.
As a control, culture supernatant was replaced by 2× McIlvaine buffer (pH 8) and treated in the same way as the sample.
Slides were rinsed twice in 0.01 M PBS (pH 7.4), transferred to 0.07 M citrate buffer (pH 6.0; Bio-Optica), and treated in a microwave oven at 750 W for 1 min for permeabilization.
Tissue samples of both origins were paraffin-embedded, cut to 3 5 µm sections, heated at 80°C for 1 hour, dewaxed with xylene, rehydrated and treated in citrate buffer, pH 6.0, for 4 min in a pressure cooker to demask antigenic sites.
For input samples, 40 µL of chromatin solution was combined with 60 µl elution buffer and 70 µL of TE and treated in the same manner as IP samples to reverse cross links.
5 µm-depth sections were deparaffinised, rehydrated and treated in sodium citrate buffer (pH 6.0).
For immunohistochemistry, 5 μm paraffin sections were deparaffinized, rehydrated, and treated in preheated citrate buffer pH 6.0 (Dako) at 95 99 °C for 20 min in a microwave oven.
In the case of biofilms grown on graphite electrodes and treated in chlorine-free M56 buffer with 50 μg/mL ampicillin and 83 μA/currentrentherere was even a slight decrease in killing.
The slides were briefly incubated in 0.2 mol l−1 HCl for 20 min, washed with Wash Buffer, incubated for 30 min at 80°C with Pretreatment Solution (NaSCN), washed with Wash Buffer and finally treated in a Protease I solution (0.5 mg ml−1 protease buffer; pH 2) for 10 12 min at 37°C.
