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Transient expression assays in Nicotiana benthamiana were performed using the protocol reported previously (Li, X. 2011).
In addition, all patients in both groups were managed by tight glucose control, using the protocol reported by Van den Berghe [32],[32].
The AROS model was subjected to molecular dynamics simulations by using the protocol reported in the Methods.
The model obtained for AROS was subjected for 15 ns to molecular dynamics simulations using the protocol reported above to assess its conformational stability.
In situ hybridization was performed using the protocol reported by Martinez and co-workers [ 82] with several modifications.
In addition, all patients in both groups were managed by tight glucose control, using the protocol reported by Van den Berghe [ 32],[ 33].
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The oligomer identification has been performed using the protocols reported in [40] and [43].
Using primers and the protocol reported by García-Álvarez et al. (3 ), we detected a mecA variant (mecALGA251) in both isolates.
PCR amplifications were accomplished using the protocol previously reported in [34].
Cortical neurons were cultured from C57BL/6 J mice using a method modified from the protocol reported by Redmond et al. Briefly, cerebral cortices were removed from 16 18-day-old embryos, stripped of meninges and blood vessels and minced.
For EBP50 stable overexpression, MDA-MB-231 cells were transfected with pBK-CMV-HA-EBP50 plasmid or pBK-CMV-HA vector, respectively, using FuGENE6 (Roche, IN) following the protocol reported previously (Konno et al. 2009).
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