Cell - 8 September 2016

B

35S:YFP-SIM

sim/smr1

25 μm 25 μm

35S:YFP-SIM

cpr5/sim/smr1

1 cm 1 cm

35S:YFP-SIM

cpr5/sim/smr1

cpr5/sim/smr1 sim/smr1 sim/smr1

cpr5

WT

A

0 0.5 1.0 1.5

Relative BiFC intensity

CPR5-SIM

G120D-SIM

Figure S6. CKIs Release and NPC Transport Activity Change Are Both Required for ETI Activation, Related toFigure 6
(A) The G120D mutation compromises CPR5 interaction with SIM. BiFC was performed by transiently coexpressing SIM-nYFP and cYFP-CPR5/G120D in
N. benthamiana. The BiFC intensity was normalized using averaged expression levels of YFP-CPR5/G120D measured in separate experiments. Data were
plotted as relative values to the average BiFC intensity of CPR5-SIM and are presented as mean±SDM (n = 15 cells for each BiFC combination).
(B) Three-week-old WT,cpr5,sim smr1andcpr5 sim smr1plants together with isogenic lines constitutively expressing YFP-SIM insim smr1, andcpr5 sim smr1
backgrounds (upper panel). Overexpression of SIM in thesim smr1background caused serrated leaf morphology without PCD, but in thecpr5 sim smr1
background led to thecpr5-like dwarfism and PCD (lower panel). Nuclear YFP-SIM is pseudo-colored in green and chlorophyll autofluorescence is in red. Ar-
rowheads indicate enlarged nuclei as a result of SIM overactivation (lower panel).