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Plates were dried in a safety cabinet for 20 min before inoculation with bacteria.
To prove that the modulation in the inflammatory response (Figure 5) is mediated by A2AR, we treated animals with an A2AR antagonist (30 min before inoculation, ZM241385, 1 mg/kg).
WEB2170 (5 mg/Kg; i.p). was given 30 min before inoculation of EAT cells.
WEB2170 (5 mg/Kg; i.p). was given 30 min before inoculation of EAT cells and DTIC (40 μg/animal; i.p). was given every 3 days after tumour implantation.
Anaerobiosis was reached by sparging the fermentor medium with N2 gas for 5 to 10 min before inoculation and constantly stirred at 150 rpm during cultivation.
Mice were also either treated with WEB2170 or PBS (as a control) 30 min before inoculation of the thymocytes to investigate the possible involvement of PAF-R in the potentiating effect of apoptotic cells.
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In order to assess whether the increases in cell numbers in BALF were dependent on enzymatic activity per se, HYAL was heated and denatured at 60° or 95°C for 60 min, respectively, before inoculation.
All the media were autoclaved at 121°C for 20 min before the inoculation.
All the media were bubbled through nitrogen for 10 min to remove oxygen and autoclaved at 121°C for 20 min before fermentation inoculation.
Where indicated, mice were injected i.p. with the CXCR2 antagonist SB225002 at 10 mg/kg 30 min before Salmonella inoculation (Alves-Filho et al, 2010).
To diminish the risk for bacterial infection, the inocula were preheated for 10 min at 70°C before inoculation.
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