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. 2018 May 29;9:672. doi: 10.3389/fpls.2018.00672

FIGURE 5.

FIGURE 5

Quantification of protein tyrosine nitration (A) and protease activity (D) in leaves of resistant potato at the site of avr P. infestans inoculation (PCD zone) and in the surrounding area (PCD distal zone). Analyses were performed at 1, 3, 6, 24, 48, 72, and 96 h after challenge inoculation. Values represent the mean ± SD of at least three independent experiments (n = 9). Asterisks indicate values that differ significantly from mock inoculated (control) potato leaves at ∗P < 0.05. (B) Trypan blue staining illustrating the PCD zone (1) and PCD distal zone (2) at 96 hpi. (C) Representative immunoblot showing immunoreactive bands containing subtilisin-like proteins undergoing tyrosine nitration in potato leaves challenge inoculated with avr P. infestans at 96 hpi. Western blot was probed with a rabbit anti-nitrotyrosine polyclonal antibody at a 1:1.000 dilution. The loading control was determined by staining the blot with CBB R-250. The indicated band was isolated from PVDF membrane and identified by LC-MS-MS/MS as described in Supplementary Data Sheet S1.