For the nucleus subcellular colocalization, the proteins were co-infiltrated with cultures (OD600 0.1) expressing the nuclear localization signal of SV40 large T antigen fused to the RFP, kindly provided by Dr. José Navarro IBMCP, Valencia, Spain.
As described by Leastro et al. (2018) (link) for callose staining, N. benthamiana leaves were infiltrated with aniline blue (Merck KGaA, Darmstadt, Germany) solution at 0.005% concentration in sodium phosphate buffer, 70 mM, pH 9.0. The leaves were infiltrated and kept in a dark room for 2 h before confocal visualization.
As described by Leastro et al. (2018) (link) for callose staining, N. benthamiana leaves were infiltrated with aniline blue (Merck KGaA, Darmstadt, Germany) solution at 0.005% concentration in sodium phosphate buffer, 70 mM, pH 9.0. The leaves were infiltrated and kept in a dark room for 2 h before confocal visualization.
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