Cell - 8 September 2016

Figure S3. Overexpression of CPR5-C Interferes with the Function of the Endogenous CPR5 Protein, Related toFigure 3
(A) CPR5-C did not complement thecpr5mutant phenotypes. Two independent transgenic lines are shown. WT, thecpr5mutant and the complemented line
expressing the full-length CPR5 are shown as control. Expression levels of transgenes are shown in the right panel.
(B) CPR5-C co-localizes with CPR5. YFP-CPR5-C and mCherry-CPR5 were transiently co-expressed inN. benthamianaand epidermal cells were imaged 24 hr
afterAgrobacteriuminfiltration. The arrowhead indicates the nucleus (N).
(C and D) Transient expression of HA-CPR5-C but not HA-CPR5-N or HA-CPR5 induced PCD inN. benthamiana(C). PCD induced by transient expression of
YFP-CPR5-C could be suppressed by coexpression of YFP-CPR5 but not YFP-CPR5-N (D, left) at comparable protein levels (D, right). All constructs are driven by
the 35S promoter. Representative leaves were photographed at 36 hr afterAgrobacteriuminfiltration.
(E) Segregating T2 progeny of35S:YFP-CPR5-CtransgenicArabidopsislines in WT background. The presence of the transgene was determined by PCR. Two
independent lines (#3 and #4) are shown andcpr5-1mutant served as a control.
(F) qRT-PCR analysis of SA-dependent defense gene expression in35S:YFP-CPR5-Ctransgenic lines. The transcript levels of the endogenous CPR5 were not
affected in the35S:YFP-CPR5-Clines (right). Data are presented as mean±SDM (n = 3 biological replicates).
(G) Inducible expression of CPR5-C is innocuous in thecpr5mutant. IsogenicDex:YFP-CPR5-Clines in WT and thecpr5mutant background were imaged 5 days
after dexamethasone treatment (upper panel). Relative expression levels of most induced defense genes in thecpr5mutant were measured 3 days after
dexamethasone treatment using qRT-PCR (lower panel). Data are presented as mean±SDM (n = 2 biological replicates).
(H) Time course qRT-PCR analysis of SA-responsive genes inDex:YFP-CPR5-Ctransgenic plants (WT background) after dexamethasone application. Relative
expression levels of SA responsive (PR1) and transport (EDS5) genes were measured. A transgenic line with the empty vector and thecpr5mutant served as
controls. Data are presented as mean±SDM (n = 2 technical replicates). Similar results were obtained in two independent experiments.