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The presence of Econea® within the microscaffolds was confirmed by Fourier transformed infrared spectroscopy (FTIR), thermogravimetric analyses and scanning electron microscopy (SEM-EDX) and it was found to be nearly constant over the time when those are immersed in artificial seawater, while under mild agitation.
The coating suspension was kept under agitation during the coating experiments while being fed with a peristaltic pump (Provitec, DM7900, São Paulo, Brazil).
Specifically, we observed that while GFP-Rac1 localized to the plasma membrane in exponentially growing cells under agitation (Figure 1A, left panel), this fusion protein accumulated in an intracellular compartment in the absence of agitation (Figure 1A, middle panel).
The culture was incubated at 30 °C under agitation for 24 h.
The suspension was maintained under agitation at 75 °C and nitrogen bubbling for 30 min prior to add the other components.
First, 25 mL Luria-Bertani (LB) medium, supplemented with 100 μg/mL ampicillin, were grown overnight at 37 °C under agitation.
In the other experiments, measurements were carried out under agitation.
Flasks were kept at 28 °C under agitation at 120 g for 3 days.
The mixture was incubated overnight at 50 °C under agitation at 250 rpm.
After stabilization, 6.486 g of 37% formaldehyde solution was added slowly under agitation.
These bio-colloids were incubated under agitation at 27 °C for 3 or 12 h.
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